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Literature summary extracted from

  • Sriskanda, V.; Moyer, R.W.; Shuman, S.
    NAD+-dependent DNA ligase encoded by a eukaryotic virus (2001), J. Biol. Chem., 276, 36100-36109.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.5.1.2 native and mutant enzymes in Escherichia coli BL21 (DE3) Amsacta moorei entomopoxvirus

Protein Variants

EC Number Protein Variants Comment Organism
6.5.1.2 D117A no nick-ligation activity Amsacta moorei entomopoxvirus
6.5.1.2 D27A no effect on activity Amsacta moorei entomopoxvirus
6.5.1.2 D48A reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick Amsacta moorei entomopoxvirus
6.5.1.2 D52A reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick Amsacta moorei entomopoxvirus
6.5.1.2 DEL1-70 inactive in overall ligation reaction, fully functional with preadenylated nick Amsacta moorei entomopoxvirus
6.5.1.2 DEL216-532 inactive in overall ligation reaction, formation of adenylate-ligase intermediate not affected Amsacta moorei entomopoxvirus
6.5.1.2 K115A no nick-ligation activity Amsacta moorei entomopoxvirus
6.5.1.2 Y39A reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick Amsacta moorei entomopoxvirus
6.5.1.2 Y40A reduced adenylate-transfer activity, no effect on DNA-ligation with preadenylated nick Amsacta moorei entomopoxvirus
6.5.1.2 Y51A adenylate-transfer activity abolished, no effect on DNA-ligation with preadenylated nick Amsacta moorei entomopoxvirus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.5.1.2 0.009
-
 NAD+
-
 Amsacta moorei entomopoxvirus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.5.1.2 Co2+ can partially substitute for Mg2+ Amsacta moorei entomopoxvirus
6.5.1.2 Mg2+ highest activity at 5 mM Amsacta moorei entomopoxvirus
6.5.1.2 Mn2+ can partially substitute for Mg2+ Amsacta moorei entomopoxvirus
6.5.1.2 additional information Ca2+, Zn2+ and Cu2+ have no effect on activity Amsacta moorei entomopoxvirus
6.5.1.2 additional information no zinc binding motif as known from most bacterial DNA-ligases Amsacta moorei entomopoxvirus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.5.1.2 62000
-
 SDS-PAGE, recombinant enzyme Amsacta moorei entomopoxvirus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.5.1.2 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

EC Number Organism UniProt Comment Textmining
6.5.1.2 Amsacta moorei entomopoxvirus
-
 entomopoxvirus
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.5.1.2 recombinant enzymes using His-tag Amsacta moorei entomopoxvirus

Reaction

EC Number Reaction Comment Organism Reaction ID
6.5.1.2 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
a

Storage Stability

EC Number Storage Stability Organism
6.5.1.2 -80°C Amsacta moorei entomopoxvirus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.5.1.2 NAD+ + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Amsacta moorei entomopoxvirus AMP + nicotinamide nucleotide + (deoxyribonucleotide)n+m
-
 ?
6.5.1.2 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

EC Number Subunits Comment Organism
6.5.1.2 monomer zonal velocity sedimentation centrifugation, protein is found to be asymmetrically shaped Amsacta moorei entomopoxvirus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.5.1.2 6.5 9
-
 Amsacta moorei entomopoxvirus

Cofactor

EC Number Cofactor Comment Organism Structure
6.5.1.2 NAD+ required, highest activity 0.005-0.1 mM Amsacta moorei entomopoxvirus