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Literature summary for 6.5.1.1 extracted from

  • Sriskanda, V.; Kelman, Z.; Hurwitz, J.; Shuman, S.
    Characterization of an ATP-dependent DNA ligase from the thermophilic archaeon Methanobacterium thermoautotrophicum (2000), Nucleic Acids Res., 28, 2221-2228.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.5.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Methanothermobacter thermautotrophicus

Protein Variants

Protein Variants Comment Organism
D253A mutant DNA ligase reacts with ATP to form the covalent intermediate, but is unable to catalyze the full ligation reaction Methanothermobacter thermautotrophicus
K251A no DNA ligase-adenylate formation and no nick-joining Methanothermobacter thermautotrophicus

Inhibitors

Inhibitors Comment Organism Structure
ammonium sulfate 10 mM, 60% inhibition Methanothermobacter thermautotrophicus
KCl 50 mM, 70% inhibition Methanothermobacter thermautotrophicus
NaCl 50 mM, 75% inhibition, 80 mM, 90% inhibition Methanothermobacter thermautotrophicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011
-
 ATP 60°C, pH 7.5 Methanothermobacter thermautotrophicus
0.0016
-
 dATP 60°C, pH 7.5 Methanothermobacter thermautotrophicus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 10 mM, divalent cation required for activity Methanothermobacter thermautotrophicus
Mg2+ 10 mM, divalent cation required for activity Methanothermobacter thermautotrophicus
Mn2+ 10 mM, divalent cation required for activity Methanothermobacter thermautotrophicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
68000
-
 zonal velocity sedimentation Methanothermobacter thermautotrophicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m Methanothermobacter thermautotrophicus
-
 AMP + diphosphate + (deoxyribonucleotide)m+n
-
 ?

Organism

Organism UniProt Comment Textmining
Methanothermobacter thermautotrophicus Q50566
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant Mth ligase Methanothermobacter thermautotrophicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m
-
 Methanothermobacter thermautotrophicus AMP + diphosphate + (deoxyribonucleotide)m+n
-
 ?
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m strandjoining on a singly nicked DNA in the presence of a divalent cation and ATP Methanothermobacter thermautotrophicus AMP + diphosphate + (deoxyribonucleotide)m+n
-
 ?
dATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m strandjoining on a singly nicked DNA in the presence of a divalent cation and dATP, approx. 10% of activity with ATP Methanothermobacter thermautotrophicus dAMP + diphosphate + (deoxyribonucleotide)m+n
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 68000, SDS-PAGE Methanothermobacter thermautotrophicus

Synonyms

Synonyms Comment Organism
Mth ligase
-
 Methanothermobacter thermautotrophicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
-
 Methanothermobacter thermautotrophicus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
22 80 almost no activity at 22°C, approx. 25% of maximal activity at 40°C and 70°C Methanothermobacter thermautotrophicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 8.8
-
 Methanothermobacter thermautotrophicus

pH Range

pH Minimum pH Maximum Comment Organism
6.5 9 approx. 20% of maximal activity at pH 6.5, approx. 90% of maximal activity at pH 9.0 Methanothermobacter thermautotrophicus