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Literature summary for 2.5.1.17 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

Activating Compound Comment Organism Structure
KCl below 50 mM Thermoplasma acidophilum

Cloned(Commentary)

Cloned (Comment) Organism
-
Thermoplasma acidophilum
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)

Crystallization (Comment) Organism
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

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

General Stability

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

KM Value [mM]

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

Metals/Ions

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Thermoplasma acidophilum Q9HIA7
-
-
Thermoplasma acidophilum Q9HIA7 gene TA1434
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Turnover Number [1/s]

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

pH Optimum

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