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Literature summary for 6.5.1.2 extracted from

  • Timson, D.J.; Wigley, D.B.
    Functional domains of an NAD+-dependent DNA ligase (1999), J. Mol. Biol., 285, 73-83.
    View publication on PubMed
Search for more literature for 6.5.1.2

Cloned(Commentary)

Cloned (Comment) Organism
N-terminal fragment 1-318 and C-terminal fragment starting from position 397 seperately expressed in Escherichia coli BL21 Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
DELTA1-396 no self adenylation activity, DNA-binding not affected Geobacillus stearothermophilus
DELTA319-670 full self adenylation activity but strongly reduced DNA-binding and ligation activity Geobacillus stearothermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ bound to the C-terminal fragment Geobacillus stearothermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 SDS-PAGE, C-terminal fragment Geobacillus stearothermophilus
30040
-
 ESI-mass spectrometry, C-terminal fragment Geobacillus stearothermophilus
35790
-
 ESI-mass spectrometry, N-terminal fragment Geobacillus stearothermophilus
36000
-
 SDS-PAGE, N-terminal fragment Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m Geobacillus stearothermophilus DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
fragments overexpressed in Escherichia coli Geobacillus stearothermophilus

Reaction

Reaction Comment Organism Reaction ID
ATP + (deoxyribonucleotide)n-3'-hydroxyl + 5'-phospho-(deoxyribonucleotide)m = (deoxyribonucleotide)n+m + AMP + beta-nicotinamide D-nucleotide 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 Geobacillus stearothermophilus
a

Storage Stability

Storage Stability Organism
-80°C, 10% glycerol Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Geobacillus stearothermophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m DNA-repair, phosphodiester bond formation between adjacent 5'-phosphate and 3'-hydroxyl groups in double-stranded DNA Geobacillus stearothermophilus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?

Cofactor

Cofactor Comment Organism Structure
NAD+ required Geobacillus stearothermophilus