Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.5.1.2 extracted from

  • Sriskanda, V.; Schwer, B.; Ho, C.K.; Shuman, S.
    Mutational analysis of Escherichia coli DNA ligase identifies amino acids required for nick-ligation in vitro and for in vivo complementation of the growth of yeast cells deleted for CDC9 and LIG4 (1999), Nucleic Acids Res., 27, 3953-3963.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.5.1.2

Cloned(Commentary)

Cloned (Comment) Organism
capable of complementing DNA-ligase-deficient yeast, wild type and mutants expressed in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
C408A inactive, unable to complement DNA-ligase-deficient yeast Escherichia coli
C411A inactive, unable to complement DNA-ligase-deficient yeast Escherichia coli
C426A no effect in vivo, 7% of wild type activity Escherichia coli
C432A inactive, unable to complement DNA-ligase-deficient yeast Escherichia coli
D117A inactive in nick-joining, unable to complement DNA-ligase-deficient yeast Escherichia coli
DELTA1-38 inactive, unable to complement DNA-ligase-deficient yeast Escherichia coli
DELTA1-78 inactive, unable to complement DNA-ligase-deficient yeast Escherichia coli
E113A no effect in vivo, increased Km for NAD+, 40% of wild type activity Escherichia coli
E319A no effect in vivo, 69% of wild type activity Escherichia coli
K115A inactive in nick-joining, unable to complement DNA-ligase-deficient yeast Escherichia coli
K314A [DV1] accumulation of the DNA-adenylate intermediate, loss of function in vivo, strongly reduced activity Escherichia coli
Q318A no effect in vivo, 80% of wild type activity Escherichia coli
Y225A no effect in vivo, increased Km for NAD+, 25% of wild type activity Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.003
-
 NAD+ wild type enzyme Escherichia coli
0.045
-
 NAD+ E113A mutant enzyme Escherichia coli
0.076
-
 NAD+ Y225A mutant enzyme Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ zinc binding motiv, C408, C411, C432 and C426 Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
76000
-
 SDS-PAGE, recombinant protein Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m Escherichia coli 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
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
His-tag used for purification of recombinant enzymes Escherichia coli

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 Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Escherichia coli 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 Escherichia coli AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?

Cofactor

Cofactor Comment Organism Structure
NAD+ required Escherichia coli