Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
K126A | inactive | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NAD+ | inhibitory above 0.1 mM, activity is abolished at 0.5 mM | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 25% of the activity with Mg2+ | Escherichia coli | |
Mg2+ | 1-10 mM, required | Escherichia coli | |
Mn2+ | 1-10 mM, required | Escherichia coli | |
Zn2+ | no effect | Escherichia coli |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
second NAD+-dependent ligase LigB, high sequence homology to LigA, lacks zinc binding motif that is found in LigA | - |
Purification (Comment) | Organism |
---|---|
recombinant protein using His-tag | Escherichia coli |
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 |
Storage Stability | Organism |
---|---|
-80°C | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + nicked DNA | - |
Escherichia coli | AMP + diphosphate + ? | - |
? | |
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 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0008 | - |
nicked DNA | - |
Escherichia coli |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 9 | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | required, highest activity 0.007-0.06 mM | Escherichia coli |