Information on EC 6.5.1.2 - DNA ligase (NAD+)

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The expected taxonomic range for this enzyme is: dsDNA viruses, no RNA stage, Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY
6.5.1.2
-
RECOMMENDED NAME
GeneOntology No.
DNA ligase (NAD+)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
mechanism, the initial step is most likely a nucleophilic attack of the epsilon-amino group of a Lys on the adenylyl phosphorus of NAD+
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
mechanism, the initial step is most likely a nucleophilic attack of the epsilon-amino group of a Lys on the adenylyl phosphorus of NAD+; the second step is the transfer of the AMP moiety from the enzyme to the 5'-phosphoryl of the DNA substrate, recreating a diphosphate linkage and thereby preserving the high-energy bond. The third step is the nucleophilic attack of the adjacent 3'-hydroxyl group to form a phosphodiester bond and eliminate AMP
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
Q9RBF2
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m = AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
3 steps of reaction: 1. adenylation of the ligase in the presence of NAD+, 2. transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate, 3. sealing the nick through the formation of a phosphodiester bond
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
esterification
-
-
phosphodiester
-
SYSTEMATIC NAME
IUBMB Comments
Poly(deoxyribonucleotide):poly(deoxyribonucleotide) ligase (AMP-forming, NMN-forming)
Catalyses the formation of a phosphodiester at the site of a single-strand break in duplex DNA. RNA can also act as substrate, to some extent.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Deoxyribonucleate ligase
-
-
-
-
Deoxyribonucleic acid joinase
-
-
-
-
Deoxyribonucleic acid ligase
-
-
-
-
Deoxyribonucleic joinase
-
-
-
-
Deoxyribonucleic ligase
-
-
-
-
Deoxyribonucleic repair enzyme
-
-
-
-
Deoxyribonucleic-joining enzyme
-
-
-
-
DNA joinase
-
-
-
-
DNA ligase
-
-
-
-
DNA ligase
-
-
DNA ligase
-
-
DNA ligase (NAD)
-
-
-
-
DNA repair enzyme
-
-
-
-
DNA-joining enzyme
-
-
-
-
LigA
P15042
-
LigA
Escherichia coli GR501, Escherichia coli MG1655
-
-
-
LigA
Staphylococcus aureus RN4220
-
-
-
LigA
Thermotoga maritima ATCC43589
-
-
-
LigA
Q9ZHI0
-
Ligase, polynucleotide (nicotinamide adenine dinucleotide)
-
-
-
-
MimiLIG
-
-
MsEPV DNA ligase
-
-
NA1 ligase
Q2Q452
-
NAD(+)-dependent DNA ligase
-
-
NAD+ dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
Q837V6
-
NAD+-dependent DNA ligase
P15042
-
NAD+-dependent DNA ligase
Escherichia coli MG1655
-
-
-
NAD+-dependent DNA ligase
O87703
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
P63973
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
Q9AIU7
-
NAD+-dependent DNA ligase
Staphylococcus aureus RN4220
-
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
Thermotoga maritima ATCC43589
-
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
-
-
NAD+-dependent DNA ligase
Q9ZHI0
-
NAD+-dependent DNA ligase
-
-
Polydeoxyribonucleotide synthase (NAD+)
-
-
-
-
Polydeoxyribonucleotide synthase [NAD+]
-
-
-
-
Polynucleotide ligase
-
-
-
-
Polynucleotide synthetase
-
-
-
-
Polynucleotide synthetase (nicotinamide adenine dinucleotide)
-
-
-
-
Synthetase, polydeoxyribonucleotide (nicotinamide adenine dinucleotide)
-
-
-
-
T4 DNA ligase
-
-
Taq DNA ligase
-
-
Tfi DNA ligase
-
-
-
-
Tfu DNA ligase
Q9HH07
-
TNA1_lig
Q2Q452
-
Tth DNA ligase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
37259-52-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
one NAD+ and one ATP dependent DNA-ligase, ATP dependent enzyme displays limited activity in nick sealing
-
-
Manually annotated by BRENDA team
one NAD+ dependent and two ATP dependent DNA-ligases, only NAD+ dependent enzyme is essential for growth
-
-
Manually annotated by BRENDA team
Borrelia hermsii DAH
-
-
-
Manually annotated by BRENDA team
second NAD+-dependent ligase LigB, high sequence homology to LigA, lacks zinc binding motif that is found in LigA
-
-
Manually annotated by BRENDA team
strain 595 (lamdagt4 lop11 lig+ S7) with 500fold higher activity than the wild type strain
-
-
Manually annotated by BRENDA team
strain GR501
-
-
Manually annotated by BRENDA team
strain K12
-
-
Manually annotated by BRENDA team
Escherichia coli GR501
strain GR501
-
-
Manually annotated by BRENDA team
Escherichia coli K12
strain K12
-
-
Manually annotated by BRENDA team
Escherichia coli MG1655
-
-
-
Manually annotated by BRENDA team
entomopoxvirus
-
-
Manually annotated by BRENDA team
Staphylococcus aureus RN4220
-
-
-
Manually annotated by BRENDA team
strain A3(2)
-
-
Manually annotated by BRENDA team
strain NA1
SwissProt
Manually annotated by BRENDA team
Thermotoga maritima ATCC43589
-
-
-
Manually annotated by BRENDA team
strain TAK16D
-
-
Manually annotated by BRENDA team
Thermus sp. TAK16D
strain TAK16D
-
-
Manually annotated by BRENDA team
HB8, wild-type and site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
DNA ligase enhances efficiency of DNA polymerase in long PCR amplification
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + diphosphate + (deoxyribonucleotide)n+m
show the reaction diagram
Q9HH07, -
Tfu DNA ligase displays nick joining and blunt-end ligation activity using either ATP or NAD+ as cofactor. Tfu DNA ligase is likely to use the same catalytic residue with the two cofactors. Higher affinity towards NAD+ than towards ATP
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9ZHI0, -
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9RBF2
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9F150
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
O87703
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
joining of the 5'-phosphoryl of a DNA chain to the 3'-hydroxyl of an RNA molecule in two ways: 1. Oligo(A) is joined to the 5'-phosphoryl of oligo(dA) on a poly(dT) template strand. 2. the enzyme can circularize poly[d(A-T)]pU in which UMP occupies the 3'-hydroxyl terminal position
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
(dT)8, (dT)10, (dT)12-18
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
joining of a single-strand-scission in double-stranded DNA
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
blunt-end ligation, intermolecular joining to yield linear oligomers, but no circular DNA forms in presence of high concentrations of polyethylene glycols
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
blunt-end ligation in presence of macromolecular solutes, inactive in absence
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
oligonucleotides as short as six or seven in length can be joined if annealed to long complementary deoxyribonucleotides, joining of 5'-phosphoryl terminus of DNA chain to the 3'-hydroxyl terminus of RNA
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA ligase requires two DNA termini, the 5'-terminus, carrying a phosphate group, and the 3'-terminus, a hydroxyl group. These termini must reside on a double-stranded molecule, DNA:DNA or DNA:RNA. Both strands of the duplex may terminate, in the form of a staggered end or a blunt end, and the ligase then requires a second similar double-stranded terminus to join the two in an intermolecular reaction. Alternatively, the two termini may be provided by a nick in just one strand of a duplex, which the enzyme will then seal
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
cohesive-end DNA restriction fragments
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
bacteriophage T5 DNA, which contains single strand interruptions at defined positions in one of the complementary strands
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
formation of linear oligomers by intermolecular cohesive-end ligation
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
the self-complementary polymer, poly(dA-dT), forms a looped-back structure that DNA ligase can join to yield a circular molecule
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
joins oligo(dT) that is base-paired to poly(dA) or oligo(dA) base-paired to poly(dT)
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
PvuII endonuclease-digested pBR322 DNA, formation of linear oligomers ranging from dimer through pentamer
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
formation of linear oligomers by intermolecular blunt-end ligation in presence of high concentrations of polymer polyethylene glycol
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
nick-closing activity
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
nick-closing activity
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
P49421, -
nick-closing activity
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
nick-closing activity
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
catalyzes the joining of polynucleotide strands provided they have juxtaposed 3'-hydroxyl and 5'-phosphoryl end groups aligned in a duplex structure: e.g. annealed ends of lamdda DNA, endogenous nicks in T5 DNA, interruptions created by the action of pancreatic DNAse, annealed fragments generated by the staggered cutting action of some restriction endonucleases
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
no ligation of blunt-ended or flush-ended DNAs
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-, O66880
activity depends on the base pairs on both sides of the nick, T/A
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9ZHI0, -
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-, O66880
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9RBF2
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9F150
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
LigA is essential for growth of Mycobacterium tuberculosis
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
acridinium-ester labelled DNA
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
cohesive ends of restriction endonuclease digested lambda-DNA
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9HH07, -
Tfu DNA ligase displays nick joining and blunt-end ligation activity using either ATP or NAD+ as cofactor. Tfu DNA ligase is likely to use the same catalytic residue with the two cofactors. Higher affinity towards NAD+ than towards ATP
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
the enzyme seals sticky ends efficiently, but has little activity on 1-nucleotide gap or blunt-ended DNA substrates even in presence of polyethylene glycol. MsEPV DNA ligase readily joins DNA nicks with mismatches at either side of the nick junction, except for mismatches at the nick junction containing an A base in the template strand (A/A, G/A, and C/A). The enzyme can join DNA probes on RNA templates. MsEPV DNA ligase joins mismatches at the 3' side of the nick more frequently than at the 5' side of the nick on an RNA template. No activity with ATP
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
although NAD+-dependent DNA ligase is essential for mycobacterial viability, only low levels of protein are required for growth
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q2Q452
enzyme is active with ATP and with NAD+
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
in addition to the unique N-terminal domain Ia that stimulates ligase-AMP formation, there are three C-terminal domains that extend from the OB domain: a small zinc-binding (Zn) domain, a helix-hairpin-helix domain, and a BRCA1 C-terminal domain
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
Hind III digested plasmid
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Thermotoga maritima ATCC43589
-
Hind III digested plasmid
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Mycoplasma pneumoniae M29, Escherichia coli MG1655
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Escherichia coli K12
-
-, DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Thermus sp. TAK16D
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Staphylococcus aureus RN4220
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
DNA ligase plays the key role in DNA replication of joining the nascent small pieces of DNA at the replication fork. The enzyme participates in the synthesis and repair of DNA
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
the enzyme is indispensable for normal cell growth and inviability of mutants seems to be primarily the result of an inability to seal Okazaki fragments
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
joining of short DNA fragments formed during DNA replication and so enabling DNA synthesis to progress in an overall 3'-5' direction on the antiparallel strand of the double helix, while continual 5'-3' synthesis proceeds on the other strand. Plays a role during genetic recombination and in the repair of UV-damaged DNA
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
the enzyme plays a pivotal role in DNA replication, repair and recombination
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9AIU7, -
-
-
-
?
NAD+ + 5'-d(ACCATTCCTGATTCTAAGTG)-3' + 5'-d(CTCAGGTCGACAGTCTGCGG)-3'
?
show the reaction diagram
-
100% ligation efficiency
-
-
?
NAD+ + 5'-d(GCCATTCCTGATTCTAAGTG)-3' + 5'-d(CTCAGGTCGACAGTCTGCGG)-3'
?
show the reaction diagram
-
100% ligation efficiency
-
-
?
NADH + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
NADH has a significantly higher Km as NAD+
-
-
-
Thionicotinamide derivative of NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
significantly higher Km as NAD+
-
-
-
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + diphosphate + (deoxyribonucleotide)n+m
show the reaction diagram
-
NAD+-dependent DNA ligase LigN is non-essential for cell viability. Haloferax volcanii also encodes the ATP-dependent DNA ligase LigA. As with LigN, LigA is also non-essential for cell viability. Simultaneous inactivation of both proteins is lethal, however, indicating that they share an essential function
-
-
?
additional information
?
-
-
NAD+/nicotinamide nucleotide exchange reaction
-
-
-
additional information
?
-
-
no activity with NADH, NADP, NADPH, ATP and AMP
-
-
-
additional information
?
-
Thermotoga maritima, Thermotoga maritima ATCC43589
-
purified DNA ligase exhibits activity on DNA fragments with cohesive termini, and no activity is detected on blunt-end DNA. DNA ligase promotes the amplification of long DNA fragments from the genomic DNA of Thermotoga maritima
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + diphosphate + (deoxyribonucleotide)n+m
show the reaction diagram
-
NAD+-dependent DNA ligase LigN is non-essential for cell viability. Haloferax volcanii also encodes the ATP-dependent DNA ligase LigA. As with LigN, LigA is also non-essential for cell viability. Simultaneous inactivation of both proteins is lethal, however, indicating that they share an essential function
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9ZHI0, -
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-, O66880
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9RBF2
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Q9F150
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
LigA is essential for growth of Mycobacterium tuberculosis
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
although NAD+-dependent DNA ligase is essential for mycobacterial viability, only low levels of protein are required for growth
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
-
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
DNA ligase plays the key role in DNA replication of joining the nascent small pieces of DNA at the replication fork. The enzyme participates in the synthesis and repair of DNA
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
the enzyme is indispensable for normal cell growth and inviability of mutants seems to be primarily the result of an inability to seal Okazaki fragments
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
joining of short DNA fragments formed during DNA replication and so enabling DNA synthesis to progress in an overall 3'-5' direction on the antiparallel strand of the double helix, while continual 5'-3' synthesis proceeds on the other strand. Plays a role during genetic recombination and in the repair of UV-damaged DNA
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
?
show the reaction diagram
-
the enzyme plays a pivotal role in DNA replication, repair and recombination
-
-
-
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + beta-nicotinamide D-ribonucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
-
-
-
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Escherichia coli K12
-
DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA
-
?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
show the reaction diagram
Thermus sp. TAK16D
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NAD+
-
required, highest activity at 0.03 mM
NAD+
-
required, highest activity 0.005-0.1 mM
NAD+
-
required, binding of cofactor induces conformational rearangement within the active site
NAD+
-, O66880
required, highest activity at 0.5 mM
NAD+
-
required, highest activity 0.007-0.06 mM
NAD+
Q9HH07, -
Tfu DNA ligase displays nick joining and blunt-end ligation activity using either ATP or NAD+ as cofactor. Tfu DNA ligase is likely to use the same catalytic residue with the two cofactors. Higher affinity towards NAD+ than towards ATP
NAD+
-
dependent on
NAD+
-
NAD+ presence is essential for exhibiting DNA ligase activity
NAD+
-
required, optimal activity at 1 mM NAD+
NAD+
-
in the presence of NAD+, Escherichia coli DNA ligase is activated
ATP
Q9HH07, -
Tfu DNA ligase displays nick joining and blunt-end ligation activity using either ATP or NAD+ as cofactor. Tfu DNA ligase is likely to use the same catalytic residue with the two cofactors. Higher affinity towards NAD+ than towards ATP
additional information
-
no activity with ATP
-
additional information
-
does not require ATP as cofactor
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
60% as active as Mg2+ in activation as reported in one study, no activity in another
Ca2+
Q9F150
40% of activity obtained with Mg2+
Ca2+
-, O66880
supports formation of DNA-adenylate intermediate
Ca2+
-
activates wild-type enzyme and mutant enzyme DELTA582-667
Ca2+
-
10fold lower activation than Mn2+, maximal activity at 2 mM
Ca2+
-
in presence of Ca2+ MtuLigA is able to carry out the first two steps of ligation reaction that is the transfer of AMP to enzyme from NAD+ and further transfer of it to substrate to form DNA-adenylate intermediate. Ca2+ can not support nick closure activity
Ca2+
-
the enzyme requires a divalent cation like Ca2+, optimal activity at about 20 mM Ca2+
Co2+
-
can partially substitute for Mg2+
Co2+
-
25% of the activity with Mg2+
Cu2+
Q9F150
30% of activity obtained with Mg2+
K+
-
stimulates at low concentrations
K+
-
stimulates
K+
-
activates at 1-150 mM
K+
Q9RBF2
25 mM, stimulates activity
K+
-
highest activity below 10 mM
KCl
-
stimulation by 300 mM KCl, in 10% w/v polyethylene glycol 6000 solution or 150-300 mM KCl in 15% polyethylene glycol 6000 solution. Stimulation of intermolecular blunt end ligation by 150-250 mM KCl in 15% polyethylene glycol 600 solution
KCl
-
no activity is detected in the absence of KCl. Maximum activity at 3.2 M KCl, close to the intracellular KCl concentration of Haloferax volcanii cells
Mg2+
-
optimal concentration: 1-3 mM; requires divalent cations, Mn2+ or Mg2+
Mg2+
-
requires divalent cations, Mn2+ or Mg2+
Mg2+
-
optimal concentration: 5-9 mM; requires divalent cations, Mn2+ or Mg2+
Mg2+
-
required
Mg2+
Q9RBF2
required
Mg2+
Q9F150
required
Mg2+
-
highest activity at 5 mM
Mg2+
-, O66880
5 mM
Mg2+
-
1-10 mM, required
Mg2+
-
or Mn2+ required, maximal activity at 10-15 mM
Mg2+
-
best metal-activator of wild-type enzyme and mutant enzyme DELTA582-667
Mg2+
Q2Q452
activates
Mg2+
-
10fold lower activation than Mn2+, maximal activity at 1 mM
Mg2+
-
divalent cation required, optimal concentration: 10 mM
Mg2+
-
Mg2+ activates at 4.0 mM
Mg2+
-
optimum concentration at 5 mM
Mg2+
-
the enzyme requires a divalent cation like Mg2+, optimal activity at about 20 mM Mg2+
Mg2+
-
required for activity, maximal activity occurs with 10-20 mM MgCl2 in the presence of 30 mM each of KCl and (NH4)2SO4
MgCl2
Q9HH07, -
optimum concentration: 2 mM
Mn2+
-
requires divalent cations, Mn2+ or Mg2+
Mn2+
-
activation at 0.2-1.0 mM, inhibition at higher concentration; requires divalent cations, Mn2+ or Mg2+
Mn2+
-
requires divalent cations, Mn2+ or Mg2+
Mn2+
-
optimal concentration: 3-6 mM; requires divalent cations, Mn2+ or Mg2+
Mn2+
Q9F150
50% of activity obtained with Mg2+
Mn2+
-
can partially substitute for Mg2+
Mn2+
-, O66880
5 mM, most effective metal cofactor
Mn2+
-
1-10 mM, required
Mn2+
-
or Mg2+ required, maximal activity at 1 mM
Mn2+
-
activates wild-type enzyme and mutant enzyme DELTA582-667
Mn2+
-
preference for Mn2+ as cofactor, maximal activity at 1 mM
Na+
-
inhibitory
NaCl
-
stimulation of intermolecular cohesive-end ligation by 200 mM NaCl, in 10% w/v polyethylene glycol 6000 solution, or 100-200 mM NaCl in 15% polyethylene glycol 6000 solution. Stimulation of intermolecular blunt end ligation by 100-150 mM NaCl in 15% polyethylene glycol 600 solution
NH4+
-
stimulates at low concentrations
NH4+
-
stimulates
NH4+
-
activates at 1-150 mM
NH4+
Q9RBF2
10 mM, stimulates activity
NH4+
-
highest activity below 10 mM
Ni2+
Q9F150
10% of activity obtained with Mg2+
Ni2+
-, O66880
supports formation of DNA-adenylate intermediate
Rb+
-
stimulates
Zn2+
-
slight activation
Zn2+
-
zinc binding motif, Cys437, Cys440, Cys455, Cys460
Zn2+
-
1 mol per mol of enzyme, Cys406, Cys409, Cys422 and Cys427 form a zinc binding motif
Zn2+
Q9RBF2
0.24 mol per mol protein
Zn2+
Q9F150
5% of activity obtained with Mg2+
Zn2+
-
bound to the C-terminal fragment
Zn2+
-
zinc binding motiv, C408, C411, C432 and C426
Zn2+
-
no effect
Zn2+
Q2Q452
activates
Mn2+
-
the enzyme requires a divalent cation like Mn2+, maximal activity at 3 mM Mn2+
additional information
-
no zinc-binding motif as known for bacterial DNA-ligase
additional information
-
Ca2+, Zn2+ and Cu2+ have no effect on activity; no zinc binding motif as known from most bacterial DNA-ligases
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(29S,34S,36R,37R)-1,1'-(1,3-phenylene)bis(3-((S)-1-((3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)-3-hydroxypropyl)urea)
-
-
(29S,34S,36R,37R)-1,1'-(1,4-phenylene)bis(3-((S)-1-((3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)-3-ethoxy-3-oxopropyl)urea)
-
-
(3S,3'R)-3,3'-(propane-1,3-diyldiimino)bis[3-[(3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl]propan-1-ol]
-
-
(4R)-N4-(6-chloro-2-methoxyacridin-9-yl)-N1,N1-diethylpentane-1,4-diamine
-
-
(8R,10R)-10-((4S,5S,6S)-4-Amino-5-hydroxy-6-methyl-tetrahydro-pyran-2-yloxy)-6,8,11-trihydroxy-8-hydroxymethoxymethyl-1-methoxy-7,8,9,10-tetrahydro-naphthacene-5,12-dione
-
IC50: 0.005 mM
(NH4)2SO4
-
(NH4)2SO4 concentrations higher than 30 mM are inhibitory. At 100 mM KCl, concentrations of (NH4)2SO4 above 12 mM are inhibitory
1,N6-etheno-NAD+
-
-
12-(6-piperidin-1-ylhexyl)-7,12-dihydro-6H-[1]benzothiepino[5,4-b]indole
-
-
2,2'-([4-[(6-chloro-2-methoxyacridin-9-yl)amino]pentyl]imino)diethanol
-
-
2,4-diamino-5-oxo-5H-chromeno[2,3-b]pyridine-3-carbonitrile
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
2,4-diamino-7-dimethylamino-pyrimido[4,5-d]pyrimidine
-
IC50 of 0.0001 mg/ml, specific inhibitor of NAD+-dependent DNA ligase, inhibits competitively with respect to NAD+ and specifically inhibits enzyme adenylation, but not DNA adenylation or ligation
2-amino-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
2-amino-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
2-amino-7,8-difluoro-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations, worse inhibitor than 2-amino-7-fluoro-5-oxo-4a,10a-dihydro-5H-chromeno[2,3-b]pyridine-3-carboxamide
2-amino-7-fluoro-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations, better inhibitor than 2-amino-7,8-difluoro-5-oxo-4a,10a-dihydro-5H-chromeno[2,3-b]pyridine-3-carboxamide
2-methylthio ADP
-
-
2-methylthio-ATP
-
-
2-methylthio-ATP
-
potent inhibitor
3,3'-(decane-1,10-diylbis(azanediyl))bis(3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propan-1-ol)
-
-
3,3'-(propane-1,3-diylbis(azanediyl))bis(3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[2,3-d][1,3]dioxol-5-yl)propan-1-ol)
-
-
3-((R)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-3-(3-phenoxymethyl-thioureido)-propionic acid ethyl ester
-
IC50: 0.004 mM
3-((R)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-3-[3-(3-{3-[1-((S)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-2-ethoxycarbonyl-ethyl]-ureido}-phenyl)-ureido]-propionic acid ethyl ester
-
IC50: 0.0277 mM
3-amino-6-[(6-chloro-2-methoxyacridin-9-yl)amino]heptan-1-ol
-
-
3-chloro-9-[[5-(diethylammonio)pentan-2-yl]amino]-7-methoxyacridin-2-aminium
-
-
3-chloro-N9-[5-(diethylamino)pentan-2-yl]-7-methoxyacridine-2,9-diamine
-
-
5-[[[(4-chlorophenyl)amino]carbonyl](phenylmethyl)amino]-5,6-dideoxy-1,2-O-(1-methylethylidene)-3-O-(phenylmethyl)-alpha-L-gluco-heptofuranuronic acid ethyl ester
-
IC50: 0.00965 mM
6-chloro-2-methoxy-N-[4-(piperidin-1-yl)pentyl]acridin-9-amine
-
-
6-chloro-9-[[5-(diethylammonio)pentan-2-yl]amino]-2-methoxyacridinium
-
-
ADP
-
inhibits adenylation reaction
AMP
-
inhibits adenylation reaction
ATP
-
inhibits adenylation reaction
cAMP
-
inhibits adenylation reaction
Chloroquine
-
; effective inhibitor
Chloroquine diphosphate
-
IC50: 0.0014 mM
Doxorubicin
-
IC50: 0.0033 mM
Doxorubicin
-
doxorubicin inhibits the ATP-dependent DNA ligase of bacteriophage T4 and the Escherichia coli NAD+-dependent ligase with a similar potency
ethyl 3-(1-benzyl-3-(4-chlorophenyl)ureido)-3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propanoate
-
-
ethyl 3-(3-benzoylthioureido)-3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propanoate
-
-
ethyl-3-O-benzyl-5,6-dideoxy-1,2-O-(1-methylethylidene)-5-[[(phenylcarbonyl)carbamoyl]amino]-alpha-L-xylo-heptofuranuronate
-
-
ethyl-3-O-benzyl-5-[benzyl[(4-chlorophenyl)carbamoyl]amino]-5,6-dideoxy-1,2-O-(1-methylethylidene)-alpha-L-xylo-heptofuranuronate
-
-
hydroxychloroquine
-
specific inhibitor of the Escherichia coli enzyme
hydroxychloroquine
-
-
KCl
-
inhibition by KCl occurs at greater than 30 mM (NH4)2SO
methotrexate
-
IC50 of 0.008 mg/ml
Mn2+
-
activation at 0.2-1.0 mM, inhibition at higher concentration
N'-(6-chloro-2-methoxyacridin-9-yl)-N,N-diethylbutane-1,4-diamine
-
-
N'-(6-chloro-2-methoxyacridin-9-yl)-N,N-dimethylbutane-1,4-diamine
-
-
N,N'-bis[4-((4-(diethylamino)-1-methylbutyl)amino)-quinoline-6-yl]sebacamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
N,N'-bis[4-((4-(diethylamino)-1-methylbutyl)amino)-quinoline-8-yl]adipamide
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
N,N'-bis[[(3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl]methyl]dodecane-1,12-diamine
-
-
N,N-bis(4-aminobutyl)-N'-(6-chloro-2-methoxyacridin-9-yl)butane-1,4-diamine
-
-
N,N-bis(4-aminobutyl)-N'-(6-chloro-2-methoxyacridin-9-yl)pentane-1,5-diamine
-
-
N-(4-aminobutyl)-N-(3-aminopropyl)-N'-(6-chloro-2-methoxyacridin-9-yl)butane-1,4-diamine
-
-
N-(4-aminobutyl)-N-(3-aminopropyl)-N'-(6-chloro-2-methoxyacridin-9-yl)pentane-1,5-diamine
-
-
N-(4-aminobutyl)-N-[3-[(6-chloro-2-methoxyacridin-9-yl)amino]propyl]butane-1,4-diamine
-
-
N-(6-chloro-2-methoxyacridin-9-yl)butane-1,4-diamine
-
-
N-[6-piperidin-1-ylhexyl]-6,7-dihydrobenzo[b]indolo[2,3-d]thiepine
-
LX competes with NAD+ andf inhibits the enzyme with IC50 in the low micromolar range, LX exhibits around 15fold better inhibition of the Mycobacterium tuberculosis enzyme compared to the human DNA ligase
N1,N1-bis(2-chloroethyl)-N4-(6-chloro-2-methoxyacridin-9-yl)pentane-1,4-diamine
-
-
N1-(7-chloro-[4]quinolyl)-N4,N4-bis-(2-hydroxy-ethyl)-1-methyl-butanediyldiamine
-
IC50: 0.046 mM
N4-(7-chloro-7,8-dihydroquinolin-4-yl)-N1,N1-diethylpentane-1,4-diamine
-
-
N4-(7-chloroquinolin-4-yl)-N1,N1-diethylpentame-1,4-diamine
-
-
Na+
-
above 20-150 mM
NAD+
-
inhibitory above 0.1 mM, activity is abolished at 0.5 mM
pyridochromanone
-
IC50: 0.0006 mM
pyridochromanones
-
differently substituted compounds tested and all found to be inhibitory, competitive with NAD+, inhibitory in nano-molar rage without affecting human DNA-ligase (ATP-depentend) activity
-
Quinacrine
-
powerful inhibitor, exhibits good inhibition at nanomolar concentrations
additional information
-
not inhibited by trimethoprim
-
additional information
-
not inhibited by cinchonidine, quinine, and N,N'-bis[4chloroquinolin-8-yl]succinamide
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(NH4)2SO4
-
with 30 mM (NH4)2SO4, the activity is maximal
ATP
Q2Q452
activates
Bovine serum albumin
Q9RBF2
0.025 mg/ml, stimulating
-
dithiothreitol
Q9RBF2
10 mM, stimulating
KCl
Q9F150
highest activity at 150 mM
KCl
-, O66880
150 mM
KCl
-
activation by KCl is observed at (NH4)2SO4 concentrations below 30 mM, in the absence of (NH4)2SO4 ligase activity is activated by KCl over 0-100 mM, and at 100 mM KCl, further activation occurs at up to 12 mM (NH4)2SO4
NaCl
Q9F150
highest activity at 200 mM
NAD+
Q2Q452
activates
NH4Cl
Q9F150
highest activity at 200 mM
Polyamines
-
blunt-end ligation are linear oligomers. Optimal concentration: caldopentamine, 0.05 mM, thermine, 0.1-0.2 mM, spermine, 0.2 mM, thermospermine, 0.4 mM, sperminediol, 0.75 mM
-
Polyethylene glycol
-
in presence of high molecular weight polyethylene glycol 20000, 6000 or 1000, 8-28%, the enzyme catalyzes blunt-end intermolecular joining to yield linear oligomers, but no circular DNA forms. In presence of low molecular polyethylene glycols 400, 200, 8-80%, or the monomer ethylene glycol, 16-80%, the circular forms formed by intramolecular ligation are also detected
potassium glutamate
Q9F150
highest activity at 350 mM
Macromolecular solutes
-
blunt-end ligation in presence of high concentrations of macromolecular solutes, inactive in absence, e.g. polyethylene glycol 6000, bovine plasma albumin, or Ficoll 70
-
additional information
-
no requirement for sulfhydryl reagent
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.14
-
(dT)10
-
-
2.5e-05
0.00056
5'-phosphate terminus of deoxyribonucleotides
-
-
-
2.5e-05
-
5'-phosphoryl termini of poly(dA-dT)
-
-
-
2.5e-06
-
acridinium-ester labelled DNA
-
pH 7.4, 21C
-
1.16
-
ATP
Q9HH07, -
pH 6.5, 55C
0.000136
-
DNA
-
pH 7.6, 21C, C/G matched nick DNA substrate
1.85e-05
-
NAD+
-
-
0.00275
-
NAD+
-
pH 7.4, 21C
0.003
-
NAD+
-
wild type enzyme
0.0058
-
NAD+
-
pH 7.6, 21C
0.045
-
NAD+
-
E113A mutant enzyme
0.076
-
NAD+
-
Y225A mutant enzyme
0.69
-
NAD+
Q9HH07, -
pH 6.5, 55C
8e-05
-
nicked DNA
-, O66880
pH 7.6, 65C
-
0.00017
-
nicked DNA
Q9RBF2
4C, pH 7.8
-
0.00018
-
nicked DNA
-
18C, pH 7.8
-
0.00024
-
nicked DNA
-
45C, pH 7.8
-
0.0003
-
nicked DNA
Q9RBF2
18C, pH 7.8
-
0.00047
-
nicked DNA
-
60C, pH 7.8
-
0.00063
-
nicked DNA
Q9RBF2
25C, pH 7.8
-
0.0007
-
nicked DNA
-
30C, pH 7.8
-
0.00204
-
nicked DNA
-
45C, pH 7.8
-
0.105
-
nicked DNA
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.4
-
DNA
-
pH 7.6, 21C, C/G matched nick DNA substrate
0.1
-
NAD+
-
pH 8.0, 21C
0.0008
-
nicked DNA
-
-
-
0.01
-
nicked DNA
Q9RBF2
4C, pH 7.8
-
0.016
-
nicked DNA
-
45C, pH 7.8
-
0.021
-
nicked DNA
-
18C, pH 7.8
-
0.03
-
nicked DNA
-
30C, pH 7.8
-
0.034
-
nicked DNA
Q9RBF2
18C, pH 7.8
-
0.047
-
nicked DNA
Q9RBF2
25C, pH 7.8
-
0.053
-
nicked DNA
-
45C, pH 7.8
-
0.132
-
nicked DNA
-
60C, pH 7.8
-
0.033
-
nicked substrate
-, O66880
pH 7.6, 65C
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0014
-
2,4-diamino-7-dimethylamino-pyrimido[4,5-d]pyrimidine
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.085
-
(29S,34S,36R,37R)-1,1'-(1,3-phenylene)bis(3-((S)-1-((3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)-3-hydroxypropyl)urea)
-
-
0.225
-
(29S,34S,36R,37R)-1,1'-(1,4-phenylene)bis(3-((S)-1-((3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)-3-ethoxy-3-oxopropyl)urea)
-
-
0.0462
-
(3S,3'R)-3,3'-(propane-1,3-diyldiimino)bis[3-[(3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl]propan-1-ol]
-
-
0.005
-
(8R,10R)-10-((4S,5S,6S)-4-Amino-5-hydroxy-6-methyl-tetrahydro-pyran-2-yloxy)-6,8,11-trihydroxy-8-hydroxymethoxymethyl-1-methoxy-7,8,9,10-tetrahydro-naphthacene-5,12-dione
-
IC50: 0.005 mM
0.12
-
(Rp)-3',5'-cyclic adenosine monophosphothiolate
-
37C, pH 7.4, DNA ligation
0.054
-
1,N6-etheno-NAD+
-
pH and temperature not specified in the publication
0.0135
-
12-(6-piperidin-1-ylhexyl)-7,12-dihydro-6H-[1]benzothiepino[5,4-b]indole
-
-
0.0009
-
2,4-diamino-5-oxo-5H-chromeno[2,3-b]pyridine-3-carbonitrile
-
-
7.5e-05
-
2-(butylsulfanyl)adenosine
-
pH 7.5, 22C
8.1e-05
-
2-(butylsulfanyl)adenosine
-
pH 7.5, 22C
0.000136
-
2-(butylsulfanyl)adenosine
-
pH 7.5, 22C
0.000507
-
2-(butylsulfanyl)adenosine
-
pH 7.5, 22C
0.00127
-
2-(butylsulfanyl)adenosine
-
pH 7.5, 22C
2.1e-05
-
2-(cyclobutylmethoxy)-5'-deoxy-5'-fluoroadenosine
-
pH 7.5, 22C
5.2e-05
-
2-(cyclobutylmethoxy)-5'-deoxy-5'-fluoroadenosine
-
pH 7.5, 22C
6.3e-05
-
2-(cyclobutylmethoxy)-5'-deoxy-5'-fluoroadenosine
-
pH 7.5, 22C
7.6e-05
-
2-(cyclobutylmethoxy)-5'-deoxy-5'-fluoroadenosine
-
pH 7.5, 22C
0.000158
-
2-(cyclobutylmethoxy)-5'-deoxy-5'-fluoroadenosine
-
pH 7.5, 22C
1.9e-05
-
2-(cyclopentyloxy)-5'-deoxyadenosine
-
pH 7.5, 22C
7.6e-05
-
2-(cyclopentyloxy)-5'-deoxyadenosine
-
pH 7.5, 22C
0.000112
-
2-(cyclopentyloxy)-5'-deoxyadenosine
-
pH 7.5, 22C
0.000117
-
2-(cyclopentyloxy)-5'-deoxyadenosine
-
pH 7.5, 22C
0.00037
-
2-(cyclopentyloxy)-5'-deoxyadenosine
-
pH 7.5, 22C
0.0001
-
2-amino-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
-
8e-05
-
2-amino-7,8-difluoro-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
-
4e-05
-
2-amino-7-fluoro-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxamide
-
-
0.0041
-
2-methylthio ADP
-
37C, pH 7.4, DNA ligation
0.0011
-
2-methylthio ATP
-
37C, pH 7.4, DNA ligation
0.0005
-
2-methylthio-ATP
-
-
0.26
-
3,3'-(decane-1,10-diylbis(azanediyl))bis(3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propan-1-ol)
-
-
0.0462
-
3,3'-(propane-1,3-diylbis(azanediyl))bis(3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[2,3-d][1,3]dioxol-5-yl)propan-1-ol)
-
-
0.004
-
3-((R)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-3-(3-phenoxymethyl-thioureido)-propionic acid ethyl ester
-
IC50: 0.004 mM
0.0277
-
3-((R)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-3-[3-(3-{3-[1-((S)-6-benzyloxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-yl)-2-ethoxycarbonyl-ethyl]-ureido}-phenyl)-ureido]-propionic acid ethyl ester
-
IC50: 0.0277 mM
3.8e-05
-
5'-deoxy-2-(spiro[2.2]pent-1-ylmethoxy)adenosine
-
pH 7.5, 22C
6e-05
-
5'-deoxy-2-(spiro[2.2]pent-1-ylmethoxy)adenosine
-
pH 7.5, 22C
6.2e-05
-
5'-deoxy-2-(spiro[2.2]pent-1-ylmethoxy)adenosine
-
pH 7.5, 22C
8.2e-05
-
5'-deoxy-2-(spiro[2.2]pent-1-ylmethoxy)adenosine
-
pH 7.5, 22C
0.00018
-
5'-deoxy-2-(spiro[2.2]pent-1-ylmethoxy)adenosine
-
pH 7.5, 22C
1e-05
-
5'-deoxy-5'-fluoro-2-[(trans-4-methylcyclohexyl)oxy]adenosine
-
IC50 less 0.00001 mM, pH 7.5, 22C
3.9e-05
-
5'-deoxy-5'-fluoro-2-[(trans-4-methylcyclohexyl)oxy]adenosine
-
pH 7.5, 22C
0.000106
-
5'-deoxy-5'-fluoro-2-[(trans-4-methylcyclohexyl)oxy]adenosine
-
pH 7.5, 22C
0.000158
-
5'-deoxy-5'-fluoro-2-[(trans-4-methylcyclohexyl)oxy]adenosine
-
pH 7.5, 22C
0.00032
-
5'-deoxy-5'-fluoro-2-[(trans-4-methylcyclohexyl)oxy]adenosine
-
pH 7.5, 22C
0.00965
-
5-[[[(4-chlorophenyl)amino]carbonyl](phenylmethyl)amino]-5,6-dideoxy-1,2-O-(1-methylethylidene)-3-O-(phenylmethyl)-alpha-L-gluco-heptofuranuronic acid ethyl ester
-
IC50: 0.00965 mM
0.046
-
Chloroquine
-
chloroquine inhibits the enzyme with an IC50 of about 0.046 mM
0.053
-
Chloroquine
-
-
0.0014
-
Chloroquine diphosphate
-
IC50: 0.0014 mM
0.0033
-
Doxorubicin
-
IC50: 0.0033 mM
0.00965
-
ethyl 3-(1-benzyl-3-(4-chlorophenyl)ureido)-3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propanoate
-
-
0.004
-
ethyl 3-(3-benzoylthioureido)-3-(6-(benzyloxy)-2,2-dimethyl-dihydro-5H-furo[3,2-d][1,3]dioxol-5-yl)propanoate
-
-
0.004
-
ethyl-3-O-benzyl-5,6-dideoxy-1,2-O-(1-methylethylidene)-5-[[(phenylcarbonyl)carbamoyl]amino]-alpha-L-xylo-heptofuranuronate
-
-
0.00965
-
ethyl-3-O-benzyl-5-[benzyl[(4-chlorophenyl)carbamoyl]amino]-5,6-dideoxy-1,2-O-(1-methylethylidene)-alpha-L-xylo-heptofuranuronate
-
-
0.063
-
hydroxychloroquine
-
-
0.0018
-
N,N'-bis[4-((4-(diethylamino)-1-methylbutyl)amino)-quinoline-6-yl]sebacamide
-
-
0.009
-
N,N'-bis[4-((4-(diethylamino)-1-methylbutyl)amino)-quinoline-8-yl]adipamide
-
-
0.0114
-
N,N'-bis[[(3aR,6S,6aR)-6-(benzyloxy)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl]methyl]dodecane-1,12-diamine
-
-
0.046
-
N1-(7-chloro-[4]quinolyl)-N4,N4-bis-(2-hydroxy-ethyl)-1-methyl-butanediyldiamine
-
IC50: 0.046 mM
0.046
-
N4-(7-chloro-7,8-dihydroquinolin-4-yl)-N1,N1-diethylpentane-1,4-diamine
-
-
0.046
-
N4-(7-chloroquinolin-4-yl)-N1,N1-diethylpentame-1,4-diamine
-
-
0.0006
-
pyridochromanone
-
IC50: 0.0006 mM
0.0015
-
Quinacrine
-
-
0.0015
-
Sp-3',5'-cyclic adenosine monophosphothiolate
-
37C, pH 7.4, DNA ligation
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
-
-
additional information
-
-
electrochemical assay for DNA ligase activity. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of nicked DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin
additional information
-
-
non-isotopic method for the determination of activity of the thermostable NAD-dependent DNA ligase from Thermus thermophilus HB8. The assay uses the cohesive ends of restriction endonuclease digested lambda-DNA as substrate. Separation of ligated from unligated cohesive end-containing fragments via agarose gel electrophoresis, followed by digital image analysis employing a normalization procedure to correct for artifacts of gel electrophoresis, allows the accurate determination of unit ligase activity
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
-
NAD+/nicotinamide nucleotide exchange reaction
7
-
-
wild-type enzyme
7.4
7.6
-
Tris-HCl buffer
7.5
8
-
Tris-HCl buffer
8
8.6
Q9F150
-
8
-
-
sodium phosphate buffer
8
-
-
collidine/HCl buffer
8
-
-, O66880
-
8
-
-
with Hind III digested plasmid as substrate
8.5
-
-
mutant enzyme DELTA582-667
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.6
7.5
-
50% of maximal activity at pH 5.6 and 7.5, potassium phosphate buffer
6.5
7
Q9HH07, -
pH 6.5: about 80% of maximal activity, pH 7.0: optimum, pH 7.5: 10% of maximal activity
6.5
9.5
-
more than 60% of maximal activity between pH 7.5 and 8.5
7
9
Q2Q452
pH 7.0: about 50% of maximal activity, pH 9.0: about 70% of maximal activity
7.5
10.5
Q9RBF2
very low activity above or below this range
7.7
9.7
-
about 70% of maximal activity at pH 7.7-9.7
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
10
15
-
ligation of cohesive ends
20
-
-
mutant enzyme L15F has 20fold lower ligation activity in vitro than wild-type enzyme
24
37
-
cohesive-end DNA restriction fragments
27.7
-
-
p(dT)8, joining of thymidylate oligomers in the presence of poly(dA) as a template
34.5
-
-
p(dT)10, joining of thymidylate oligomers in the presence of poly(dA) as a template
37
-
-
p(dT)12-18, joining of thymidylate oligomers in the presence of poly(dA) as a template
42
-
-
mutant enzyme L15F has 60fold lower ligation activity in vitro than wild-type enzyme
55
-
P49421, -
-
60
-
-
with Hind III digested plasmid as substrate
65
72
-
nick-closing activity
65
-
-
blunt-end ligation
65
-
Q9F150
above 65C
70
-
-
mutant enzyme DELTA582-667 and wild-type enzyme, activity of mutant enzyme DELTA582-667 is 50% of wild-type activity
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
75
P49421, -
active in this range
15
75
-
active in this range
15
85
-
nick-closing activity observed in the range
20
37
-
activity at 20C is lower than at 37C
30
90
Q9F150
-
31
45
-
about 70% of maximal activity at 31C and at 45C
40
80
Q9HH07, -
about 50% of maximal activity at 40C and at 80C
50
80
-
50C: about 50% of maximal activity of wild-type enzyme and of mutant enzyme DELTA582-667, 80C: about 50% of maximal activity of wild-type enzyme and mutant enzyme DELTA582-667
75
90
Q2Q452
75C: about 85% of maximal activity, 90C: about 95% of maximal activity
PDB
SCOP
CATH
ORGANISM
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Streptococcus pneumoniae (strain P1031)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30000
-
-
SDS-PAGE, C-terminal fragment
30040
-
-
ESI-mass spectrometry, C-terminal fragment
35790
-
-
ESI-mass spectrometry, N-terminal fragment
36000
-
-
SDS-PAGE, N-terminal fragment
62000
-
-
SDS-PAGE, recombinant enzyme
72000
-
-
SDS-PAGE
72130
-
-
calculated from amino acid sequence
73610
-
-
calculated from amino acid sequence
74260
-
Q9RBF2
Q-TOF-MS-mass spectrometry, non-adenylated form
74590
-
Q9RBF2
Q-TOF-mass spectrometry, adenylated form
75080
-
Q9AIU7, -
calculated from amino acid sequence
75290
-
-
mass spectrometry
75940
-
-
calculated from DNA-sequence
76000
-
-
SDS-PAGE, recombinant protein
77000
-
-
sedimentation equilibrium ultracentrifugation
80400
-
-
sedimentation equilibrium ultracentrifugation
82000
-
Q9F150
SDS-PAGE
additional information
-
-
nucleotide sequence
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 80000, SDS-PAGE, DNA-ligase-adenylate intermediate
?
P49421, -
x * 79487, calculation from nucleotide sequence
?
Q2Q452
x * 63000, recombinant enzyme, SDS-PAGE
?
-
x * 70000, SDS-PAGE
?
-
x * 72000, SDS-PAGE
?
-
x * 78000, SDS-PAGE
?
Thermotoga maritima ATCC43589
-
x * 78000, SDS-PAGE
-
monomer
-
1 * 78500, SDS-PAGE
monomer
-
1 * 74000, SDS-PAGE of denatured and reduced enzyme
monomer
-
zonal velocity sedimentation centrifugation, protein is found to be asymmetrically shaped
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystals of adenylation domain are grown by vapor diffusion using hanging drops. Crystals belong to space group C2 with unit cell parameters of a = 90.2 A, b = 86.1 A, c = 57.1 A and beta = 101. Visualization of NAD+ recognition by an NAD+-dependent ligase
-
sitting drop vapor diffusion method at 22C. 2.3 A crystal structure of Escherichia coli LigA bound to an adenylylated nick, which captures LigA in a state poised for strand closure and reveals the basis for nick recognition. LigA envelops the DNA within a protein clamp
-
hanging drop vapour phase diffusion method, K114A mutant protein fragment
-
adenylation domain of NAD+-dependent ligase with bound AMP
-
hanging drop vapor diffusion method, using 2.1-2.5 M ammonium sulfate and 0.1 M MES or HEPES pH 6.8-7.5
Q9AIU7, -
microseeding technique
-
vapor diffusion method
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
8.8
-
the enzyme retains over 80% of its activity after holding a pH ranging from 7.2 to 8.8 for 1 h at 80C
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
activity decreases remarkably below 4C
24
-
-
stable for 1 week
37
-
-
stable for 1 week
65
-
-
2 days, 50% loss of activity
70
-
-
wild-type enzyme is stable for 40 min, about 20% loss of activity after 180 min. Mutant enzyme DELTA582-667 loses 20% of its activity after 80 min and 90% of its activity after 170 min
72
-
-
2 h, stable below, destabilized above
80
-
Q9HH07, -
pH 6.5, 3 h, stable
90
-
Q9HH07, -
pH 6.5, 15 min, 50% loss of activity
90
-
-
wild-type enzyme is stable for 20 min, about 50% loss of activity after 100 min. Mutant enzyme DELTA582-667 loses 30% of its initial activity after 40 min, completely loses activity after 80 min
91
-
P49421, -
half-life: 7 min
91
-
-
half-life: 26 min
95
-
Q9F150
60 min, less than 25% of activity lost
95
-
-
50% of activity lost within 5 min
95
-
-
50% of activity lost within 15 min
95
-
-
the purified enzyme has a half-life of over 30 min at 95C
100
-
-
the half-life of heat inactivation at 100C is about 3 min
additional information
-
-
thermal stability is increased by cofactor binding
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the guanidine hydrochloride-induced equilibrium unfolding of deadenylated and adenylated enzyme form can be described by a four-state transition model
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, 10% glycerol
-
-20C, 20 mM Tris/HCl, pH 7.1, 50 mM NaCl, 0.1 mM EDTA, 50% glycerol
Q9RBF2
4C, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant enzymes using His-tag
-
recombinant protein
Q9F150
recombinant proteins
-
His-tag used for purification of recombinant enzymes
-
large scale
-
Ni-agarose column chromatography
-
recombinant
-
recombinant protein from Escherichia coli GR501 using His-tag
-
recombinant protein using His-tag
-
fragments overexpressed in Escherichia coli
-
recombinant
-
recombinant enzyme
-
MimiLIG is produced in Escherichia coli as a His10-Smt3 fusion and purified from a soluble bacterial lysate by Ni2+-agarose chromatography
-
His10-LigA fusion
-
recombinant protein from Escherichia coli GR501 using His-tag
-
recombinant enzyme from Escherichia coli
Q9RBF2
HisTrap FF column chromatography, hydroxyapatite type I column chromatography, and HiPrep Q column chromatography
Q9AIU7, -
recombinant proteins from Escherichia coli
-
Sepharose column chromatography and hydroxyapatite column chromatography
-
recombinant protein from Escherichia coli GR501 using His-tag
-
heat treatment and immobilized metal affinity column chromatography
-
DEAE column chromatographyand phosphor-cellulose column chromatography
-
recombinant protein
-
wild-type and mutants expressed in Escherichia coli
-
DEAE column chromatography and phosphor-cellulose column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
native and mutant enzymes in Escherichia coli BL21 (DE3)
-
expressed in Escherichia coli
-, O66880
expressed in Escherichia coli
Q9F150
full length and truncated enzymes expressed in Escherichia coli BL21 (DE3)
-
expressed in Escherichia coli
-
adenylation domain
-
capable of complementing DNA-ligase-deficient yeast, wild type and mutants expressed in Escherichia coli BL21(DE3)
-
expressed in Escherichia coli BL21 (DE3)
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli GR501, that contains a temperature sensitive mutation in ligA and is therefore unable to grow above 42C, expression of protein enables Escherichia coli GR501 to grow at 44C
-
ligase fragments and mutant
-
1-318 fragment with K114A mutation
-
N-terminal fragment 1-318 and C-terminal fragment starting from position 397 seperately expressed in Escherichia coli BL21
-
expressed in Escherichia coli BL21(DE3) cells
-
wild-type and mutant forms of LigN are expressed in Escherichia coli
-
expression in Escherichia coli
-
wild-type and mutant proteins Y36A, D46A, Y49A, D50A, and K113A are produced in Escherichia coli
-
expressed in Escherichia coli GR501, that contains a temperature sensitive mutation in ligA and is therefore unable to grow above 42C, expression of protein enables Escherichia coli GR501 at 44C
-
produced in Escherichia coli as a His10-LigA fusion
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli
Q9RBF2
overexpression in Escherichia coli
P49421, -
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
wild type and fragments expressed in Escherichia coli GI724
-
expressed in Escherichia coli BL21 (DE3) cells
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli GR501, that contains a temperature sensitive mutation in ligA and is therefore unable to grow above 42C, expression of protein enables Escherichia coli GR501 at 44C
-
expression in Escherichia coli BL21(DE3)
Q9HH07, -
expression in Escherichia coli
Q2Q452
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli
-
deletion mutants, expression in Escherichia coli
-
expressed in Escherichia coli BL26
-
native and selenomethionine-substituted enzyme
-
overexpression in Escherichia coli
-
expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D117A
-
no nick-ligation activity
D27A
-
no effect on activity
D48A
-
reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick
D52A
-
reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick
DEL1-70
-
inactive in overall ligation reaction, fully functional with preadenylated nick
DEL216-532
-
inactive in overall ligation reaction, formation of adenylate-ligase intermediate not affected
K115A
-
no nick-ligation activity
Y39A
-
reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick
Y40A
-
reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick
Y51A
-
adenylate-transfer activity abolished, no effect on DNA-ligation with preadenylated nick
DELTA1-525
-
no DNA-binding
DELTA1-90
-
very low self adenylation activity, active in DNA-ligation, reduced DNA-binding
DELTA1-90,363-719
-
no self adenylation
DELTA1-90,487-719
-
very low self adenylation activity
DELTA363-719
-
full self adenylation activity
DELTA517-719
-
full self adenylation activity
DELTA636-719
-
active in DNA-ligation
K128A
-
mutant enzyme shows no ligation activity
C408A
-
inactive, unable to complement DNA-ligase-deficient yeast
C411A
-
inactive, unable to complement DNA-ligase-deficient yeast
C426A
-
no effect in vivo, 7% of wild type activity
C432A
-
inactive, unable to complement DNA-ligase-deficient yeast
D117A
-
inactive in nick-joining, unable to complement DNA-ligase-deficient yeast
D117E
-
3% of ligase activity of wild-type enzyme, lethal mutation
D117N
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
D138A
-
37% of ligase activity of wild-type enzyme
D283A
-
57% of ligase activity of wild-type enzyme
D285E
-
4% of ligase activity of wild-type enzyme, lethal mutation
D285N
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
D32A
-
traces of nick joining activity
D32E
-
traces of nick joining activity
D32N
-
9% of nick joining activity, strongly reduced self-adenylation
D36A
-
traces of nick joining activity
D36E
-
4% of nick joining activity, strongly reduced self-adenylation
D36N
-
12% of nick joining activity, strongly reduced self-adenylation
D450A
-
the helix-hairpin-helix domain mutant retains DNA relaxation function, concordant with the ability to effectively ligate nicks
D452A
-
mutant with 68% of wild type activity
D551A
-
mutant with 120% of wild type activity; the helix-hairpin-helix domain mutant retains DNA relaxation function, concordant with the ability to effectively ligate nicks
DELTA1-38
-
nick joining activity abolished, no self adenylation, active with pre-adenylated nick
DELTA1-38
-
inactive, unable to complement DNA-ligase-deficient yeast
DELTA1-592
-
fragment corresponding to BRCT domain binds to a wider range of substrates compared to full-length LigA
DELTA1-78
-
nick joining activity abolished, no self adenylation, active with pre-adenylated nick
DELTA1-78
-
inactive, unable to complement DNA-ligase-deficient yeast
DELTA593-671
-
mutant lacking the BRCT domain, 3fold reduced in vitro ligation activity and reduced DNA binding
E10A
-
90% of nick joining activity
E113A
-
no effect in vivo, increased Km for NAD+, 40% of wild type activity
E143A
-
52% of ligase activity of wild-type enzyme
E173A
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
E173D
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
E173Q
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
E319A
-
no effect in vivo, 69% of wild type activity
E519A
-
mutant with 71% of wild type activity; the helix-hairpin-helix domain mutant retains DNA relaxation function, concordant with the ability to effectively ligate nicks
G118A
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
G172A
-
36% of ligase activity of wild-type enzyme
G286A
-
14% of ligase activity of wild-type enzyme
G489A
-
the mutant retains 25% of wild type activity
G489D
-
mutant with 0.8% of wild type activity
G489V
-
mutant with 3.2% of wild type activity
G521A
-
the mutant is 50fold less active than wild type LigA
G521D
-
mutant with less than 0.1% of wild type function
G521V
-
mutant with less than 0.1% of wild type function
G553A
-
mutant with 38% of wild type activity
G553D
-
mutant with 2.6% of wild type activity
G553V
-
mutant with 0.9% of wild type activity
H23A
-
10% of nick joining activity
H23Y
-
88% of nick joining activity
I384A
-
the mutant is defective for nick sealing, retaining 4% of wild type activity
K115A
-
inactive in nick-joining, unable to complement DNA-ligase-deficient yeast
K115Q
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
K115R
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
K126A
-
inactive
K290A
-
13% of ligase activity of wild-type enzyme, lethal mutation
K314A [DV1]
-
accumulation of the DNA-adenylate intermediate, loss of function in vivo, strongly reduced activity
K314Q
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
K314R
-
5% of ligase activity of wild-type enzyme, lethal mutation
K627A
-
mutant with 40% of wild type activity
K634A/K635A
-
mutant with 44% of wild type activity
K648A
-
mutant with 37% of wild type activity
K651A
-
mutant with 35% of wild type activity
L119A
-
110% of ligase activity of wild-type enzyme
L15F
-
at 20C mutant enzyme L15F has 20fold lower ligation activity in vitro, and its activity is reduced at 42C, resulting in 60fold lower ligation activity than wild-type LigA
N198A
-
26% of ligase activity of wild-type enzyme
N355A
-
mutant with 30% of wild type activity
Q318A
-
no effect in vivo, 80% of wild type activity
Q330A
-
the mutation has no apparent effect on nick sealing acivity
Q386A
-
mutant with 30% of wild type activity
R200A
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
R200K
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
R200Q
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
R208A
-
2% of ligase activity of wild-type enzyme, lethal mutation
R208K
-
1.2% of ligase activity of wild-type enzyme, lethal mutation
R208Q
-
less than 0.1% of ligase activity of wild-type enzyme, lethal mutation
R277A
-
1.1% of ligase activity of wild-type enzyme, lethal mutation; rate of isolated sealing step is 20% of the activity of wild-type enzyme
R277K
-
5.6% of ligase activity of wild-type enzyme, themperature-sensitive mutation, rate of isolated sealing step is 4% of the activity of wild-type enzyme
R277Q
-
1.8% of ligase activity of wild-type enzyme, lethal mutation, rate of isolated sealing step is 10% of the activity of wild-type enzyme
R333A
-
the oligonucleotide-binding domain mutation strongly suppresses DNA relaxation
R379A
-
the oligonucleotide-binding domain mutation strongly suppresses DNA relaxation
R487A
-
mutant with 6% of wild type activity
R510A
-
the helix-hairpin-helix domain mutant retains DNA relaxation function, concordant with the ability to effectively ligate nicks
R614A
-
mutant with 14% of wild type activity
V288A
-
74% of ligase activity of wild-type enzyme
V383A
-
the mutant is defective for nick sealing, retaining 4% of wild type activity
Y225A
-
no effect in vivo, increased Km for NAD+, 25% of wild type activity
Y22A
-
traces of nick joining activity, strongly reduced self-adenylation
Y22S
-
9% of nick joining activity
Y35A
-
2% of nick joining activity, strongly reduced self-adenylation
Y35S
-
23% of nick joining activity
L15F
Escherichia coli GR501
-
at 20C mutant enzyme L15F has 20fold lower ligation activity in vitro, and its activity is reduced at 42C, resulting in 60fold lower ligation activity than wild-type LigA
-
DELTA1-396
-
no self adenylation activity, DNA-binding not affected
DELTA319-670
-
full self adenylation activity but strongly reduced DNA-binding and ligation activity
K114A
-
prevention of partial adenylation of the enzyme
D141A
-
adenylation-defective mutant
K139A
-
adenylation-defective mutant
D46A
-
mutant protein displayes only trace sealing activity
D50A
-
nick sealing activity is 6% of wild-type activity
Y36A
-
mutant protein displayes only trace sealing activity
DELTA1-390
-
DNA binding, but no self adenylation activity
DELTA582-667
-
lower thermal stability than wild-type enzyme
D286E
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
G287A
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme. The G287A mutation has a major effect on the second step
K291R
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
V289I
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
A644I
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
E654A
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
G617I
-
mutation in BRCT domain, mutant enzyme does not shows any detectable ligation with nicked substrate in excess
G634I
-
mutation in BRCT domain, ligation activity is about 15% of the activity with wild-type enzyme with nicked substrate in excess
G638I
-
mutation in BRCT domain, ligation activity is 30-40% lower than that with wild-type enzyme with nicked substrate in excess
K619R
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
K640R
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
L647A
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
R606A
-
mutation in BRCT domain, ligation activity is 30-40% lower than that with wild-type enzyme with nicked substrate in excess
S623A
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
T599A
-
mutation in BRCT domain, ligation activity is 30-40% lower than that with wild-type enzyme with nicked substrate in excess
V624A
-
mutation in BRCT domain, comparable ligation activity with wild-type enzyme with nicked substrate in excess
D286E
Thermus sp. TAK16D
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
-
G287A
Thermus sp. TAK16D
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme. The G287A mutation has a major effect on the second step
-
K291R
Thermus sp. TAK16D
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
-
V289I
Thermus sp. TAK16D
-
mutant enzyme shows reduced reaction rate on both match and mismatch nicked substrates compared to wild-type enzyme
-
C412M
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C412T
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C412V
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C415A
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C415M
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C415T
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C415V
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C428T
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C433A
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C433M
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C433T
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
C433V
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120A
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120E
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120G
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120N
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120V
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
D120Y
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
G339A
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
G339D
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
G339E
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K118H
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K118L
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K118P1(CCC)
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K118P2(CCG)
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K118R
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K2942(CTC)
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Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K294L1(CTG)
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K294P
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K294Q
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K294R
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
R337E
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
R337K
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
R337Q
-
Site-directed mutants: K118R, K118H, K118L, K118P1(CCC), K118P2(CCG), D120E, D120N, D120Y, D120G, D120A, D120V, K294R, K294Q, K294L1(CTG), K2942(CTC), K294P, K294P, R337K, R337Q, R337E, G339A, G339D, G339E, C412A, C412V, C412T, C412M, C415A, C415V, C415T, C415M, C428A, C428T, C433A, C433V, C433T and C433M. Studies show that residue K118 plays an essential role in the adenylation step, residue D120 may facilitate the deadenylation step, residue G339 and C433 may be involved in formation of the phosphodiester bond
K113A
-
mutant protein is inert
additional information
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truncated version of MimiLIG (C-DELTA79), composed of residues 1-557, but lacking the C-terminal 79 amino acids that comprise the BRCT domain shows reduced specific activity in nick joining by 75fold without affecting the ligase adenylylation step. The DELTA-RCT mutant of MimiLIG is impaired in sealing at a preadenylylated nick
Y49A
-
nick sealing activity is 6% of wild-type activity
additional information
-
deletion of just the BRCT domain from MtuLigA results in total loss of activity in in vitro assays
DELTA316-667
-
active in self adenylation but unable to bind DNA
additional information
-
no ligation activity when both fragments are mixed together
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
NAD+-dependent ligases can serve as a valuable target in the development of chemotherapeutics for the treatment of numerous human ailments
analysis
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DNA ligase can be used to determine the ability of other enzymes to act at nicks and gaps in duplex DNA molecules; DNA ligase can be used to study the primary and secondary structure of DNA molecules; DNA ligase, in combination with polynucleotide kinase, can be used to identify 3'- and 5'-end groups at single-strand interruptions by nearest neighbor analysis; DNA ligase is an essential reagent in studies on nucleic acid structure and metabolism
analysis
-
high-throughput assay for the adenylation reaction of bacterial DNA ligase
medicine
-
potential target for antiobiotics
medicine
-
potential target for antibiotics
medicine
-
NAD+-dependent ligases can serve as a valuable target in the development if chemotherapeutics for the treatment of numerous human ailments
synthesis
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an important use of DNA ligase is the preparation of recombinant DNA molecules for use in the cloning of DNA; DNA ligase is an indispensible reagent in the chemical synthesis of double-stranded DNA of specific nucleotide sequence
synthesis
-
DNA ligase is used for cDNA cloning by replacement synthesis
medicine
-
potential target for antibiotics due to the fact that eukaryotic cells use ATP instead of NAD+ as cofactor for DNA-ligase
medicine
O87703
NAD+-dependent ligases can serve as a valuable target in the development of chemotherapeutics for the treatment of numerous human ailments
biotechnology
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LigN is unique amongst DNA ligase enzymes in displaying maximal DNA strand joining activity at above 3 M salt levels. As such the LigN enzyme has potential both as a novel tool for biotechnology and as a model enzyme for studying the adaptation of proteins to high intracellular salt levels
medicine
-
although NAD+-dependent DNA ligase is essential for mycobacterial viability, only low levels of protein are required for growth. very efficient inhibition of enzyme activity would be required if NAD+-dependent DNA ligase is to be useful as an antibiotic target in mycobacteria
medicine
-
NAD+-dependent ligases can serve as a valuable target in the development of chemotherapeutics for the treatment of numerous human ailments
medicine
-
potential target for antibiotics
medicine
-
NAD+-dependent ligases can serve as a valuable target in the development of chemotherapeutics for the treatment of numerous human ailments
medicine
-
potential target for antibiotics