BRENDA - Enzyme Database

The cobalamin-independent methionine synthase enzyme captured in a substrate-induced closed conformation

Ubhi, D.K.; Robertus, J.D.; J. Mol. Biol. 427, 901-909 (2015)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
2.1.1.14
analysis
development of a robust fluorescence-based activity assay suitable for high-throughput screening
Candida albicans
Crystallization (Commentary)
EC Number
Crystallization
Organism
2.1.1.14
binding of L-homocysteine or methionine results in conformational rearrangements at the amino acid binding pocket, moving the catalytic zinc into position to activate the thiol group. Active-site residues Asn126 and Tyr660 play key roles in catalysis
Candida albicans
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.14
N126A
13% of wild-type activity
Candida albicans
2.1.1.14
Y660A
no residual activity
Candida albicans
2.1.1.14
Y660F
93% of wild-type activity
Candida albicans
2.1.1.14
Y660Q
14% of wild-type activity
Candida albicans
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.025
-
L-homocysteine
37C, pH not specified in the publication
Candida albicans
2.1.1.14
0.087
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
37C, pH not specified in the publication
Candida albicans
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.14
Candida albicans
P82610
-
-
2.1.1.14
Candida albicans ATCC MYA-2876
P82610
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
734490
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
734490
Candida albicans ATCC MYA-2876
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.45
-
L-homocysteine
37C, pH not specified in the publication
Candida albicans
2.1.1.14
0.49
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
37C, pH not specified in the publication
Candida albicans
Application (protein specific)
EC Number
Application
Commentary
Organism
2.1.1.14
analysis
development of a robust fluorescence-based activity assay suitable for high-throughput screening
Candida albicans
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.1.1.14
binding of L-homocysteine or methionine results in conformational rearrangements at the amino acid binding pocket, moving the catalytic zinc into position to activate the thiol group. Active-site residues Asn126 and Tyr660 play key roles in catalysis
Candida albicans
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.14
N126A
13% of wild-type activity
Candida albicans
2.1.1.14
Y660A
no residual activity
Candida albicans
2.1.1.14
Y660F
93% of wild-type activity
Candida albicans
2.1.1.14
Y660Q
14% of wild-type activity
Candida albicans
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.025
-
L-homocysteine
37C, pH not specified in the publication
Candida albicans
2.1.1.14
0.087
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
37C, pH not specified in the publication
Candida albicans
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
734490
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
734490
Candida albicans ATCC MYA-2876
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.45
-
L-homocysteine
37C, pH not specified in the publication
Candida albicans
2.1.1.14
0.49
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
37C, pH not specified in the publication
Candida albicans