Information on EC 3.1.21.2 - deoxyribonuclease IV

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY
3.1.21.2
-
RECOMMENDED NAME
GeneOntology No.
deoxyribonuclease IV
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Endonucleolytic cleavage of ssDNA at apurinic/apyrimidinic sites to 5'-phosphooligonucleotide end-products
show the reaction diagram
-
-
-
-
Endonucleolytic cleavage of ssDNA at apurinic/apyrimidinic sites to 5'-phosphooligonucleotide end-products
show the reaction diagram
reaction mechanism
-
Endonucleolytic cleavage of ssDNA at apurinic/apyrimidinic sites to 5'-phosphooligonucleotide end-products
show the reaction diagram
reaction mechanism modelling
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of phosphoric diester
Q72KH8, -
-
hydrolysis of phosphoric ester
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
apurinic/apyrimidinic endonuclease
-
-
apurinic/apyrimidinic endonuclease
Escherichia coli AB1157
-
-
-
coliphage T4 endonuclease II
-
-
deoxriboendonuclease
-
-
-
-
DNA-adenine-transferase
-
-
-
-
E. coli endonuclease IV
-
-
-
-
EC 3.1.4.30
-
-
formerly
-
Endo IV
P0A6C1
-
endodeoxyribonuclease IV
-
-
-
-
Endonuclease II
-
-
Endonuclease II
-
-
endonuclease IV
-
-
endonuclease IV
-
-
endonuclease IV
P0A6C1
-
endonuclease IV
Escherichia coli AB1157
-
;
-
endonuclease IV
-
-
Nfo
-
gene name
Nfo
Escherichia coli AB1157
-
; gene name
-
nuclease, endodeoxyribo-oder redoxyendonuclease
-
-
-
-
Sco4631
Streptomyces coelicolor A(3)2
-
-
-
ScoA3McrA
Streptomyces coelicolor A(3)2
-
-
-
T4 endonuclease IV
-
-
-
-
TthNfo
Q72KH8
-
type IV methyl-dependent restriction endonuclease
-
-
type IV methyl-dependent restriction endonuclease
Streptomyces coelicolor A(3)2
-
-
-
endonuclease IV
Q72KH8
endonuclease IV of Thermus thermophilus possesses a 3'-5' exonuclease activity similar to that of Escherichia coli exonuclease III
additional information
-
the enzyme belongs to the GIY-YIG family of endonucleases
CAS REGISTRY NUMBER
COMMENTARY
37211-67-9
-
58591-37-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Escherichia coli phage, i.e. Coliphage T4
-
-
Manually annotated by BRENDA team
B.; phage T4 infected
-
-
Manually annotated by BRENDA team
gene nfo
-
-
Manually annotated by BRENDA team
gene nfo; wild type and nfo mutant
-
-
Manually annotated by BRENDA team
several strains
-
-
Manually annotated by BRENDA team
Escherichia coli AB1157
gene nfo
-
-
Manually annotated by BRENDA team
Escherichia coli B.
B.
-
-
Manually annotated by BRENDA team
gene sco4631
-
-
Manually annotated by BRENDA team
Streptomyces coelicolor A(3)2
gene sco4631
-
-
Manually annotated by BRENDA team
strain MSB8, thermophilic bacterium
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
phenotype with reduced DNA repair activity of enzyme-deficient mutant strain BW527, overview
physiological function
-
endonuclease IV is the main base excision repair enzyme involved in DNA damage induced by UVA radiation and stannous chloride; endonuclease IV recognizes and cleaves DNA on the 5' side of various oxidatively damaged bases via nucleotide incision repair (NIR)
physiological function
-
in the nucleotide incision repair pathway, an apurinic/apyrimidinic endonuclease incises duplex DNA 5' next to oxidatively damaged nucleotide. The multifunctional Escherichia coli endonuclease IV is involved in both base excision repair and nucleotide incision repair pathways, overview
physiological function
-
in vitro endonuclease activity of the Escherichia coli McrA homologue that specifically cleaves S-modified DNA
physiological function
Escherichia coli AB1157
-
endonuclease IV is the main base excision repair enzyme involved in DNA damage induced by UVA radiation and stannous chloride; endonuclease IV recognizes and cleaves DNA on the 5' side of various oxidatively damaged bases via nucleotide incision repair (NIR); in the nucleotide incision repair pathway, an apurinic/apyrimidinic endonuclease incises duplex DNA 5' next to oxidatively damaged nucleotide. The multifunctional Escherichia coli endonuclease IV is involved in both base excision repair and nucleotide incision repair pathways, overview
-
physiological function
Streptomyces coelicolor A(3)2
-
in vitro endonuclease activity of the Escherichia coli McrA homologue that specifically cleaves S-modified DNA
-
malfunction
Escherichia coli AB1157
-
phenotype with reduced DNA repair activity of enzyme-deficient mutant strain BW527, overview
-
additional information
-
a protruding loop containing a nuclease-associated modular domain 3 element is likely to be involved in substrate binding, as well as residues forming a separate nucleic acid binding surface adjacent to the active site. EndoII may bind its substrate inefficiently across the two sites in the dimer, offering a plausible explanation for the catalytic preponderance of single-strand nicks
additional information
-
Nfo catalyzed nucleotide incision and 3'-5' exonuclease activities are genetically coupled. modeling of nucleotide incision coupled to 3'-5' exonuclease activity preventing formation of lethal double-strand breaks when repairing bi-stranded clustered DNA damage, overview
additional information
Escherichia coli AB1157
-
Nfo catalyzed nucleotide incision and 3'-5' exonuclease activities are genetically coupled. modeling of nucleotide incision coupled to 3'-5' exonuclease activity preventing formation of lethal double-strand breaks when repairing bi-stranded clustered DNA damage, overview
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
21-mer C-Grec + H2O
?
show the reaction diagram
Escherichia coli, Escherichia coli AB1157
-
-
-
-
?
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides
-
-
ir
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
in vivo the enzyme produces mostly double-stranded fragments with a few internal nicks
-
-
ir
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides, hydrolysis of a phosphodiester bond near an alkylated base in native DNA without single breaks in the region, the enzyme makes predominantly single-strand break, and a few double-strand breaks, the ratio is 3.7:1
-
-
ir
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
the enzyme cleaves sites of DNA with conserved sequence elements to both the left and the right of the cleaved bonds comprising 16 basepairs with some variability tolerated, nicking of a single and both strands, the latter in two nicking events, substrate specificity, overview, in vitro the enzyme produces mostly single-stranded nicks
-
-
ir
double-stranded native DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides, the enzyme makes predominantly single-strand break, and a few double-strand breaks, the ratio is 3.7:1
-
-
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
preference for single-stranded DNA, denatured lambda-DNA, breaks exlusively adjacent to cytosine residues, no activity on denatured T4 DNA
about 150 nucleotides long
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
breakdown of host cell DNA occuring after T4 phage infection
-
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
the Ser176 of T4 endonuclease IV is crucial for the restricted and polarized dC-specific cleavage of single-stranded DNA implicated in restriction of dC-containing DNA in host Escherichia coli
-
-
?
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
Endo IV catalyzes specific endonucleolytic cleavage of the 5' phosphodiester bond of dC in ssDNA with an efficiency dependent on the surrounding nucleotide sequence. The enzyme preferentially targets 5'-dTdCdA-3' but tolerates various combinations of individual nucleotides flanking this trinucleotide sequence. Endo IV preferentially recognizes short nucleotide sequences containing 5'-TdCdA-3', which likely accounts for the limited digestion of ssDNA by the enzyme and may be responsible in part for the indispensability of a deficiency in denB for stable synthesis of dC-substituted T4 genomic DNA
-
-
?
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonuclease IV cleaves single-stranded (ss) DNA in a dC-specific manner. The Ser176 of T4 endonuclease IV is crucial for the restricted and polarized dC-specific cleavage of single-stranded DNA implicated in restriction of dC-containing DNA in host Escherichia coli
-
-
?
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
the enzyme cleaves single-stranded (ss) DNA in a dC-specific manner. Endo IV cleaves ssDNAs exclusively at the 5-proximal dC where a sequence comprises dC residues both at the 5 proximal and 3 proximal positions (a dCs tract-dependent cleavage). The dCs tract-dependent cleavage is efficient and occurs when a dCs tract has at least 6 bases. Some dCs tracts larger than 6 bases behave as that of 6 bases (an extended dCs tract), while some others do not. One decameric dCs tract is shown to be cleavable in a dCs tract-dependent manner, but that with 13 dCs is not. The dCs tract-dependent cleavage is enhanced by the presence of a third dC residue at least for a 6 or 7 dCs tract. In contrast to the dCs tract-dependent cleavage, a dCs tract-independent one is generally inefficient and if two modes are possible for a substrate DNA, a dCs tract-dependent mode prevails
-
-
?
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
Escherichia coli B.
-
preference for single-stranded DNA, denatured lambda-DNA, breaks exlusively adjacent to cytosine residues, no activity on denatured T4 DNA
about 150 nucleotides long
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
Escherichia coli B.
-
breakdown of host cell DNA occuring after T4 phage infection
-
ir
double-stranded nativeDNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides
-
-
ir
additional information
?
-
-
DNA substrate specificity, overview
-
-
-
additional information
?
-
-
endonuclease IV encoded by denB of bacteriophage T4 is implicated in restriction of deoxycytidine (dC)-containing DNA in the host Escherichia coli
-
-
-
additional information
?
-
-, P0A6C1
role for Endo IV to counteract DNA damage induced by the oxidative counterpart of UVB in Escherichia coli
-
-
-
additional information
?
-
-
reaction proceeds through a synchronous bimolecular (ANDN) mechanism with reaction free energy and barrier of -3.5 and 20.6 kcal/mol.In the course of the reaction the trinuclear active site of endonuclease IV undergoes dramatic local conformational changes: shifts in the mode of coordination of both substrate and first-shell ligands
-
-
-
additional information
?
-
-
a His-tagged derivative of ScoA3McrA cleaves S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Sco4631 cleaves DNA at multiple possible sites on either side of the S-modification. Double-strand cleavage occurs 16-28 nucleotides away from the phosphorothioate links
-
-
-
additional information
?
-
-
EndoII primarily catalyzes single-stranded nicking of DNA, 5 to 10fold less frequently double-stranded breaks are produced
-
-
-
additional information
?
-
-
Nfo-catalyzed nucleotide incision repair and exonuclease activities can generate a single-strand gap at the 5' side of 5,6-dihydrouracil residue
-
-
-
additional information
?
-
Streptomyces coelicolor A(3)2
-
a His-tagged derivative of ScoA3McrA cleaves S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Sco4631 cleaves DNA at multiple possible sites on either side of the S-modification. Double-strand cleavage occurs 16-28 nucleotides away from the phosphorothioate links
-
-
-
additional information
?
-
Escherichia coli AB1157
-
Nfo-catalyzed nucleotide incision repair and exonuclease activities can generate a single-strand gap at the 5' side of 5,6-dihydrouracil residue
-
-
-
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
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides
-
-
ir
double-stranded alkylated DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
in vivo the enzyme produces mostly double-stranded fragments with a few internal nicks
-
-
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
breakdown of host cell DNA occuring after T4 phage infection
-
ir
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
the Ser176 of T4 endonuclease IV is crucial for the restricted and polarized dC-specific cleavage of single-stranded DNA implicated in restriction of dC-containing DNA in host Escherichia coli
-
-
?
single-stranded DNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
Escherichia coli B.
-
breakdown of host cell DNA occuring after T4 phage infection
-
ir
double-stranded nativeDNA + H2O
5'-phosphooligonucleotides
show the reaction diagram
-
endonucleolytic cleavage of DNA into 5'-phosphooligonucleotides
-
-
ir
additional information
?
-
-
endonuclease IV encoded by denB of bacteriophage T4 is implicated in restriction of deoxycytidine (dC)-containing DNA in the host Escherichia coli
-
-
-
additional information
?
-
-, P0A6C1
role for Endo IV to counteract DNA damage induced by the oxidative counterpart of UVB in Escherichia coli
-
-
-
additional information
?
-
-
a His-tagged derivative of ScoA3McrA cleaves S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Sco4631 cleaves DNA at multiple possible sites on either side of the S-modification. Double-strand cleavage occurs 16-28 nucleotides away from the phosphorothioate links
-
-
-
additional information
?
-
-
EndoII primarily catalyzes single-stranded nicking of DNA, 5 to 10fold less frequently double-stranded breaks are produced
-
-
-
additional information
?
-
-
Nfo-catalyzed nucleotide incision repair and exonuclease activities can generate a single-strand gap at the 5' side of 5,6-dihydrouracil residue
-
-
-
additional information
?
-
Streptomyces coelicolor A(3)2
-
a His-tagged derivative of ScoA3McrA cleaves S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Sco4631 cleaves DNA at multiple possible sites on either side of the S-modification. Double-strand cleavage occurs 16-28 nucleotides away from the phosphorothioate links
-
-
-
additional information
?
-
Escherichia coli AB1157
-
Nfo-catalyzed nucleotide incision repair and exonuclease activities can generate a single-strand gap at the 5' side of 5,6-dihydrouracil residue
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
CaCl2
-
8% of the activity found compared to optimal Mg2+ concentration
Co2+
-
required for activity
CoCl2
-
can substitute for MgCl2, optimal concentration: 0.01 M, activity 27% higher than at optimal MgCl2 concentration
Mg2+
-
stimulates
Mg2+
-
absolutely required for nicking activity
Mg2+
Q72KH8, -
-
MgCl2
-
absolute requirement, no activity in absence, optimal concentration: 0.01 M
Mn2+
-
0.01 M can replace Mg2+
Mn2+
-
stimulates
Mn2+
Q72KH8, -
-
Zn2+
-
dependent on
Mn2+
-
required for activity
additional information
-
the enzyme shows no absolute requirement for divalent cations, no effect by Ca2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(NH4)2SO4
-
0.05 M
4-Chloromercuriphenylsulfonate
-
-
8-hydroxyquinoline
-
inhibits cleavage of alkylated dsDNA
diethyldithiocarbamic acid
-
-
EDTA
-
inhibits cleavage of non-alkylated dsDNA, 70% inhibition at 0.1 mM
EDTA
Q72KH8, -
in the presence of 10 mM EDTA enzymatic activity is completely eliminated
p-hydroxymercuribenzoate
-
-
potassium phosphate buffer
-
pH 8.3
KCl
-
24% inhibition at 0.1 M, 42% at 0.2 M, and 64% at 0.3 M
additional information
-
enzyme inhibition by chelating and sulfhydryl agents, but not by tRNA and caffeine, DTT or 2-mercaptoethanol protect against inhibition by sulfhydryl reagents
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
slight activation
Bovine serum albumin
-
130% activity at 1% at pH 8.0 and 37C
-
additional information
-
the enzyme is induced by phage T4, no activation by S-adenosyl-L-methionine, ATP, or both
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000025
-
21-mer C Grec
-
wild-type enzyme, pH 7.6, 37C
-
0.0000272
-
21-mer C Grec
-
mutant H69A, pH 7.6, 37C, in presence of Zn2+
-
0.000028
-
21-mer C Grec
-
mutant G149D, pH 7.6, 37C
-
additional information
-
additional information
-
kinetics
-
additional information
-
additional information
-
effects of dC-flanking sequences of the substrate on kinetic parameters of Endo IV
-
additional information
-
additional information
-
Km-values of Endo IV with with oligonucleotide substrates containing a target sequence and with various dC1-dC6 tracts and the effects of third dC residue in the tract
-
additional information
-
additional information
-
Km-values of the wild-type and S176N mutant forms of Endo IV are determined with series of 25-base oligonucleotides
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0003
-
21-mer C Grec
-
mutant H69A, pH 7.6, 37C, in presence of Zn2+
-
0.0088
-
21-mer C Grec
-
mutant G149D, pH 7.6, 37C
-
0.28
-
21-mer C Grec
-
wild-type enzyme, pH 7.6, 37C
-
additional information
-
additional information
-
effects of dC-flanking sequences of the substrate on kinetic parameters of Endo IV
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.012
-
21-mer C Grec
-
mutant H69A, pH 7.6, 37C, in presence of Zn2+
0
0.317
-
21-mer C Grec
-
mutant G149D, pH 7.6, 37C
0
8
-
21-mer C Grec
-
wild-type enzyme, pH 7.6, 37C
0
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
relative activity in several Escherichia coli wild-type and mutant strains, overview
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
Q72KH8, -
assay on the uracil-processing activity and 3'-5' exonuclease activity; DNA cleavage assay
7.6
-
-
assay at
8.4
9.2
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
38
-
-
assay at
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Geobacillus kaustophilus (strain HTA426)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
29000
-
Q72KH8, -
determined by SDS-PAGE
32400
-
Q9WYJ7
SDS-PAGE
additional information
-
-
sucrose density gradient sedimentation equilibrium analysis
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
tetramer
-
4 * 16000
additional information
-
EndoII forms a striking X-shaped tetrameric structure composed as a dimer of dimers, with a protruding hairpin domain not present in UvrC or I-TevI providing most of the dimerization and tetramerization interfaces. Monomer structure and topology, quarternary structure, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
mutant E118A of EndoII is crystallized in space group P21 with four monomers in the asymmetric unit, with a bound phosphate ion in one of the four active sites of EndoII likely mimicing the scissile phosphate in a true substrate complex. Crystallization of the selenomethionine-substituted E118A mutant, X-ray diffraction structure determination and analysis at 1.9-2.3 A resolution, molecular replacement
-
wild-type enzyme and mutant H69A, hanging-drop vapor diffusion method, mxing of 0.0015 ml of protein solution containing 10 mg/ml protein with 0.002 ml of reservoir solution containing 35% PEG 400 and 100mM HEPES, pH 5.5, 20C, 7 days, X-ray diffraction structure determination and analysis at 1.55 A resolution
-
using the sitting-drop vapor-diffusion method
Q9WYJ7
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
loss of 41% activity after 30 min, loss of 92% activity after 2 h, with addition 50% glycerol only 6% activity is lost within 2 h
45
-
-
10 min, fraction IV, loss of 10% activity
60
-
-
10 min, fraction IV, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glycerol and bovine serum albumin stabilize
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, purified enzyme, 40 mM potassium phosphate, pH 6.5, or 50 mM Tris-HCl, pH 8.0, with 0.1 mM DTT or 2-mercaptoethanol, moderately stable
-
-20C, Tris-buffer, pH 6.8, 50 mM NaCl, 0.5 mM DTT, 50% glycerol
Q72KH8, -
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
GST-Endo IV fusion protein
-
GST-Endo IV fusion proteins: wild-type and mutants W88R and S176N
-
phage T4 infected
-
using a amylose resin affinity column, the fusion protein is cleaved with thrombin to remove the MBP tag, the cleavage mixture is applied to a Q anion-exchange column
Q72KH8, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of GST-Endo IV fusion protein in Escherichia coli: wild-type and mutants W88R and S176N
-
gene sco4631, expression in Streptomyces lividans results in the loss of the genomic island that contains the dndA-E gene cluster, expression of N-terminally His6-tagged Sco4631 in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli
Q9WYJ7
into the pMal-c2x vector for expression in Escherichia coli TB1 cells
Q72KH8, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E118A
-
site-directed mutagenesis
S176N
-
mutant enzyme retains cleavage activity (17.5% of that of wild-type Endo IV), but loses the polarized and restricted cleavage of a dCs tract. Escherichia coli cells expressing the intact Endo IV mutant enzyme are viable and, in contrast to wild-type Endo IV, the mutant enzyme does not show detrimental effect on the host cells
W88R
-
mutant enzyme shows no enzymatic activity (less than 0.4% of that of wild-type Endo IV). Escherichia coli cells expressing the intact Endo IV mutant enzyme are viable and, in contrast to wild-type Endo IV, these mutant enzymes do not show detrimental effect on the host cells
G149D
-
site-directed mutagenesis, the mutant is deficient in both nucleotide incision repair and exonuclease activities
G149D
Escherichia coli AB1157
-
site-directed mutagenesis, the mutant is deficient in both nucleotide incision repair and exonuclease activities
-
H69A
Escherichia coli AB1157
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site-directed mutagenesis, the mutant is deficient in both nucleotide incision repair and exonuclease activities. The crystal structure of Nfo-H69A mutant reveals the loss of one of the active site zinc atoms and rearrangements of the catalytic site, but no gross changes in the overall enzyme conformation
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H508A
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catalytically inactive mutant
H508A
Streptomyces coelicolor A(3)2
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catalytically inactive mutant
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H69A
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site-directed mutagenesis, the mutant is deficient in both nucleotide incision repair and exonuclease activities. The crystal structure of Nfo-H69A mutant reveals the loss of one of the active site zinc atoms and rearrangements of the catalytic site, but no gross changes in the overall enzyme conformation
additional information
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analysis of enzyme activity in naturally occurring mutants of Escherichia coli compared to wild-type strain enzymes, overview