Cloned (Comment) | Organism |
---|---|
native and mutant enzymes in Escherichia coli BL21 (DE3) | Amsacta moorei entomopoxvirus |
Protein Variants | Comment | Organism |
---|---|---|
D117A | no nick-ligation activity | Amsacta moorei entomopoxvirus |
D27A | no effect on activity | Amsacta moorei entomopoxvirus |
D48A | reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick | Amsacta moorei entomopoxvirus |
D52A | reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick | Amsacta moorei entomopoxvirus |
DEL1-70 | inactive in overall ligation reaction, fully functional with preadenylated nick | Amsacta moorei entomopoxvirus |
DEL216-532 | inactive in overall ligation reaction, formation of adenylate-ligase intermediate not affected | Amsacta moorei entomopoxvirus |
K115A | no nick-ligation activity | Amsacta moorei entomopoxvirus |
Y39A | reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick | Amsacta moorei entomopoxvirus |
Y40A | reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick | Amsacta moorei entomopoxvirus |
Y51A | adenylate-transfer activity abolished, no effect on DNA-ligation with preadenylated nick | Amsacta moorei entomopoxvirus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.009 | - |
NAD+ | - |
Amsacta moorei entomopoxvirus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | can partially substitute for Mg2+ | Amsacta moorei entomopoxvirus | |
Mg2+ | highest activity at 5 mM | Amsacta moorei entomopoxvirus | |
Mn2+ | can partially substitute for Mg2+ | Amsacta moorei entomopoxvirus | |
additional information | Ca2+, Zn2+ and Cu2+ have no effect on activity | Amsacta moorei entomopoxvirus | |
additional information | no zinc binding motif as known from most bacterial DNA-ligases | Amsacta moorei entomopoxvirus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
62000 | - |
SDS-PAGE, recombinant enzyme | Amsacta moorei entomopoxvirus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m | Amsacta moorei entomopoxvirus | 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 |
---|---|---|---|
Amsacta moorei entomopoxvirus | - |
entomopoxvirus | - |
Purification (Comment) | Organism |
---|---|
recombinant enzymes using His-tag | Amsacta moorei entomopoxvirus |
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 | Amsacta moorei entomopoxvirus |
Storage Stability | Organism |
---|---|
-80°C | Amsacta moorei entomopoxvirus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m | - |
Amsacta moorei entomopoxvirus | 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 | Amsacta moorei entomopoxvirus | AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | zonal velocity sedimentation centrifugation, protein is found to be asymmetrically shaped | Amsacta moorei entomopoxvirus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 9 | - |
Amsacta moorei entomopoxvirus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | required, highest activity 0.005-0.1 mM | Amsacta moorei entomopoxvirus |