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Literature summary extracted from

  • Saridakis, V.; Yakunin, A.F.; Xu, X.; Anandakumar, P.; Pennycooke, M.; Gu, J.; Cheung, F.; Lew, J.M.; Sanishvili, N.; Joachimiak, A.; Arrowsmith, C.H.; Edwards, A.M.; Christendat, D.
    The structural basis for methylmalonic aciduria. The crystal structure of archaeal ATP:cobalamin adenosyltransferase (2004), J. Biol. Chem., 279, 23646-23653.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.5.1.17 KCl below 50 mM Thermoplasma acidophilum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.17
-
 Thermoplasma acidophilum
2.5.1.17 expression in Escherichia coli in strain BL21(DE3) of the wild-type enzyme and in strain B834(DE3) as selenomethionine-labeled enzyme Thermoplasma acidophilum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.17 purified recombinant wild-type and selenomethionine-labeled enzymes, crystal growth from 0.4 M ammonium phosphate, 4% methyl-pentanediol, 5% glycerol, at 20°C, X-ray diffraction structure determination and analysis at 1.5-1.9 A resolution Thermoplasma acidophilum

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.17 E126A inactive mutant Thermoplasma acidophilum
2.5.1.17 E126A no catalytic activity Thermoplasma acidophilum
2.5.1.17 E126A site-directed mutagenesis, inactive mutant Thermoplasma acidophilum
2.5.1.17 E126K inactive mutant Thermoplasma acidophilum
2.5.1.17 E126K no catalytic activity Thermoplasma acidophilum
2.5.1.17 E126K site-directed mutagenesis, inactive mutant Thermoplasma acidophilum
2.5.1.17 R119A inactive mutant Thermoplasma acidophilum
2.5.1.17 R119A no catalytic activity Thermoplasma acidophilum
2.5.1.17 R119A site-directed mutagenesis, inactive mutant Thermoplasma acidophilum
2.5.1.17 R119W inactive mutant Thermoplasma acidophilum
2.5.1.17 R119W no catalytic activity Thermoplasma acidophilum
2.5.1.17 R119W site-directed mutagenesis, inactive mutant Thermoplasma acidophilum
2.5.1.17 R124A reduced activity Thermoplasma acidophilum
2.5.1.17 R124A about half as active as wild-type Thermoplasma acidophilum
2.5.1.17 R124A site-directed mutagenesis, about 40% reduced activity compared to the wild-type enzyme Thermoplasma acidophilum
2.5.1.17 R124F reduced activity Thermoplasma acidophilum
2.5.1.17 R124F decreased kcat and increased KM Thermoplasma acidophilum
2.5.1.17 R124F site-directed mutagenesis, about 95% reduced activity compared to the wild-type enzyme Thermoplasma acidophilum
2.5.1.17 R124K inactive mutant Thermoplasma acidophilum
2.5.1.17 R124K no catalytic activity Thermoplasma acidophilum
2.5.1.17 R124K site-directed mutagenesis, inactive mutant Thermoplasma acidophilum
2.5.1.17 R124W inactive mutant Thermoplasma acidophilum
2.5.1.17 R124W no catalytic activity Thermoplasma acidophilum
2.5.1.17 R124W site-directed mutagenesis, inactive mutant Thermoplasma acidophilum

General Stability

EC Number General Stability Organism
2.5.1.17 2-mercaptoethanol stabilizes the enzyme during purfication Thermoplasma acidophilum
2.5.1.17 stable Thermoplasma acidophilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.17 0.003
-
 hydroxocobalamin
-
 Thermoplasma acidophilum
2.5.1.17 0.003
-
 hydroxocobalamin +/- 0.0004 Thermoplasma acidophilum
2.5.1.17 0.003
-
 Cobalamin pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.004
-
 Cobalamin pH 8.0, 70°C, mutant R124A, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.11
-
 ATP
-
 Thermoplasma acidophilum
2.5.1.17 0.11
-
 Mg2+ pH 8.0, 70°C, wild-type enzyme Thermoplasma acidophilum
2.5.1.17 0.11
-
 Mn2+ pH 8.0, 70°C, wild-type enzyme Thermoplasma acidophilum
2.5.1.17 0.11
-
 ATP +/- 0.01 Thermoplasma acidophilum
2.5.1.17 0.11
-
 ATP pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.14
-
 dATP
-
 Thermoplasma acidophilum
2.5.1.17 0.14
-
 dATP +/- 0.02 Thermoplasma acidophilum
2.5.1.17 0.14
-
 dATP pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.27
-
 Co2+ pH 8.0, 70°C, wild-type enzyme Thermoplasma acidophilum
2.5.1.17 0.33
-
 ATP pH 8.0, 70°C, mutant R124A, in presence of Mg2+ Thermoplasma acidophilum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.17 Co2+ activates, KD: 0.27 mM Thermoplasma acidophilum
2.5.1.17 Co2+ enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+, 39% of the activity with Mg2+ Thermoplasma acidophilum
2.5.1.17 Mg2+ activates, KD: 0.11 mM Thermoplasma acidophilum
2.5.1.17 Mg2+ enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+ Thermoplasma acidophilum
2.5.1.17 Mn2+ activates, KD: 0.11 mM Thermoplasma acidophilum
2.5.1.17 Mn2+ enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+, 83% of the activity with Mg2+ Thermoplasma acidophilum
2.5.1.17 additional information no activity with Ni2+ and Ca2+ Thermoplasma acidophilum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.17 ATP + cob(I)alamin Thermoplasma acidophilum
-
 triphosphate + adenosylcobalamin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.17 Thermoplasma acidophilum Q9HIA7
-
-
2.5.1.17 Thermoplasma acidophilum Q9HIA7 gene TA1434
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.17 0.034
-
 recombinant mutant R124F Thermoplasma acidophilum
2.5.1.17 0.28
-
 dATP Thermoplasma acidophilum
2.5.1.17 0.28
-
 +/- 0.01, dATP Thermoplasma acidophilum
2.5.1.17 0.385
-
 recombinant mutant R124A Thermoplasma acidophilum
2.5.1.17 0.47
-
 hydroxocobalamin Thermoplasma acidophilum
2.5.1.17 0.47
-
 +/- 0.02, hydroxocobalamin Thermoplasma acidophilum
2.5.1.17 0.61
-
 ATP Thermoplasma acidophilum
2.5.1.17 0.61
-
 +/- 0.02, ATP Thermoplasma acidophilum
2.5.1.17 0.619
-
 recombinant wild-type enzyme Thermoplasma acidophilum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.17 ATP + cob(I)alamin
-
 Thermoplasma acidophilum triphosphate + adenosylcobalamin
-
 ?
2.5.1.17 ATP + cobalamin enzyme is absolutely specific for ATP or dATP as adenosyl donors Thermoplasma acidophilum triphosphate + adenosylcobalamin
-
 ir
2.5.1.17 ATP + hydroxocobalamin
-
 Thermoplasma acidophilum ?
-
 ?
2.5.1.17 dATP + cob(I)alamin
-
 Thermoplasma acidophilum tripolyphosphate + deoxyadenosylcobalamin
-
 ?
2.5.1.17 dATP + cobalamin enzyme is absolutely specific for ATP or dATP as adenosyl donors Thermoplasma acidophilum triphosphate + deoxyadenosylcobalamin
-
 ir
2.5.1.17 additional information conserved residues are R119, R124, and E126 Thermoplasma acidophilum ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.17 More each subunit is composed of a bundle of 5 alpha-helices, the active site lies at the junction between the subunits Thermoplasma acidophilum
2.5.1.17 trimer
-
 Thermoplasma acidophilum
2.5.1.17 trimer gel filtration Thermoplasma acidophilum
2.5.1.17 trimer gel-filtration Thermoplasma acidophilum

Synonyms

EC Number Synonyms Comment Organism
2.5.1.17 ATP:cobalamin adenosyltransferase
-
 Thermoplasma acidophilum
2.5.1.17 MMAB
-
 Thermoplasma acidophilum
2.5.1.17 TA1434
-
 Thermoplasma acidophilum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.17 70
-
-
 Thermoplasma acidophilum
2.5.1.17 70
-
 assay at Thermoplasma acidophilum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.5.1.17 25 80
-
 Thermoplasma acidophilum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.17 0.11
-
 dATP
-
 Thermoplasma acidophilum
2.5.1.17 0.11
-
 dATP +/- 0.01 Thermoplasma acidophilum
2.5.1.17 0.11
-
 Cobalamin pH 8.0, 70°C, mutant R124A, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.11
-
 dATP pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.14
-
 ATP pH 8.0, 70°C, mutant R124A, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.18
-
 hydroxocobalamin
-
 Thermoplasma acidophilum
2.5.1.17 0.18
-
 hydroxocobalamin +/- 0.01 Thermoplasma acidophilum
2.5.1.17 0.18
-
 Cobalamin pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum
2.5.1.17 0.23
-
 ATP
-
 Thermoplasma acidophilum
2.5.1.17 0.23
-
 ATP +/- 0.01 Thermoplasma acidophilum
2.5.1.17 0.23
-
 ATP pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ Thermoplasma acidophilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.17 8
-
-
 Thermoplasma acidophilum
2.5.1.17 8
-
 assay at Thermoplasma acidophilum