BRENDA - Enzyme Database
show all sequences of 6.3.2.32

Glutathione synthetase homologs encode alpha-L-glutamate ligases for methanogenic coenzyme F420 and tetrahydrosarcinapterin biosyntheses

Li, H.; Xu, H.; Graham, D.E.; White, R.H.; Proc. Natl. Acad. Sci. USA 100, 9785-9790 (2003)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
dithiothreitol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
Cloned(Commentary)
Cloned (Commentary)
Organism
expression in Escherichia coli
Methanocaldococcus jannaschii
Inhibitors
Inhibitors
Commentary
Organism
Structure
alpha,beta-CH2-ATP
when added to a reaction mixture containing an equal amount of ATP, alpha,beta-CH2-ATP inhibits 30% of the activity
Methanocaldococcus jannaschii
KCl
in the presence of 0.1 M or 0.2 M KCl, activities are 40% and 15% relative to reactions without KCl
Methanocaldococcus jannaschii
additional information
no inhibition by beta,gamma-CH2-ATP
Methanocaldococcus jannaschii
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0012
-
coenzyme gamma-F420-2
pH 8.5, 50°C
Methanocaldococcus jannaschii
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
the CofF protein strictly requires a divalent metal ion for ligase activity. CofF enzyme incubated with 10 mM MgCl2 has about 33% of the activity supported by 10 mM MnCl2
Methanocaldococcus jannaschii
Mn2+
the CofF protein strictly requires a divalent metal ion for ligase activity. CofF enzyme incubated with 10 mM MgCl2 has about 33% of the activity supported by 10 mM MnCl2
Methanocaldococcus jannaschii
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
33278
-
1 * 33278, calculated from sequence
Methanocaldococcus jannaschii
35000
-
1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
37000
-
gel filtration
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
ATP + coenzyme gamma-F420-2 + L-glutamate
Methanocaldococcus jannaschii
the enzyme caps the gamma-glutamyl tail of the hydride carrier coenzyme F420. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
ADP + phosphate + coenzyme alpha-F420-3
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Methanocaldococcus jannaschii
Q58407
-
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant enzyme
Methanocaldococcus jannaschii
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
ATP + coenzyme gamma-F420-2 + L-glutamate
the enzyme caps the gamma-glutamyl tail of the hydride carrier coenzyme F420. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
ADP + phosphate + coenzyme alpha-F420-3
-
-
-
?
ATP + coenzyme gamma-F420-2 + L-glutamate
CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
ADP + phosphate + coenzyme alpha-F420-3
-
-
-
?
GTP + coenzyme gamma-F420-2 + L-glutamate
activity with GTP is 25% of the activity with ATP at 5 mM. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
GDP + phosphate + coenzyme alpha-F420-3
-
-
-
?
additional information
the phosphonate nucleotide analogs alpha,beta-CH2-ATP and beta,gamma-CH2-ATP support no ligase activity at concentrations of 5 mM. None of the following amino acids or analogs support ligase activity at 10-mM concentrations: D-glutamate, beta-glutamate, L-aspartate, L-glutamine, L-alpha-aminoadipate, or D,L-2-amino-4-phosphono-butyrate
700925
Methanocaldococcus jannaschii
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 33278, calculated from sequence; 1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
Synonyms
Synonyms
Commentary
Organism
CofF
-
Methanocaldococcus jannaschii
CofF protein
-
Methanocaldococcus jannaschii
gamma-F420-2:alpha-L-glutamate ligase
-
Methanocaldococcus jannaschii
MJ1001
-
Methanocaldococcus jannaschii
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Methanocaldococcus jannaschii
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
the heterologously expressed protein is thermostable
Methanocaldococcus jannaschii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at
Methanocaldococcus jannaschii
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
dithiothreitol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Methanocaldococcus jannaschii
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
alpha,beta-CH2-ATP
when added to a reaction mixture containing an equal amount of ATP, alpha,beta-CH2-ATP inhibits 30% of the activity
Methanocaldococcus jannaschii
KCl
in the presence of 0.1 M or 0.2 M KCl, activities are 40% and 15% relative to reactions without KCl
Methanocaldococcus jannaschii
additional information
no inhibition by beta,gamma-CH2-ATP
Methanocaldococcus jannaschii
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0012
-
coenzyme gamma-F420-2
pH 8.5, 50°C
Methanocaldococcus jannaschii
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
the CofF protein strictly requires a divalent metal ion for ligase activity. CofF enzyme incubated with 10 mM MgCl2 has about 33% of the activity supported by 10 mM MnCl2
Methanocaldococcus jannaschii
Mn2+
the CofF protein strictly requires a divalent metal ion for ligase activity. CofF enzyme incubated with 10 mM MgCl2 has about 33% of the activity supported by 10 mM MnCl2
Methanocaldococcus jannaschii
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
33278
-
1 * 33278, calculated from sequence
Methanocaldococcus jannaschii
35000
-
1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
37000
-
gel filtration
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
ATP + coenzyme gamma-F420-2 + L-glutamate
Methanocaldococcus jannaschii
the enzyme caps the gamma-glutamyl tail of the hydride carrier coenzyme F420. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
ADP + phosphate + coenzyme alpha-F420-3
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant enzyme
Methanocaldococcus jannaschii
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
ATP + coenzyme gamma-F420-2 + L-glutamate
the enzyme caps the gamma-glutamyl tail of the hydride carrier coenzyme F420. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
ADP + phosphate + coenzyme alpha-F420-3
-
-
-
?
ATP + coenzyme gamma-F420-2 + L-glutamate
CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
ADP + phosphate + coenzyme alpha-F420-3
-
-
-
?
GTP + coenzyme gamma-F420-2 + L-glutamate
activity with GTP is 25% of the activity with ATP at 5 mM. CofF specifically adds an alpha-linked glutamate to gamma-F420-2 produced by the Methanococcus jannaschii CofE protein. Coenzyme F420-2 is coenzyme F420 with two glutamic acid residue, coenzyme F420-3 is coenzyme F420 with three glutamic acid residues
700925
Methanocaldococcus jannaschii
GDP + phosphate + coenzyme alpha-F420-3
-
-
-
?
additional information
the phosphonate nucleotide analogs alpha,beta-CH2-ATP and beta,gamma-CH2-ATP support no ligase activity at concentrations of 5 mM. None of the following amino acids or analogs support ligase activity at 10-mM concentrations: D-glutamate, beta-glutamate, L-aspartate, L-glutamine, L-alpha-aminoadipate, or D,L-2-amino-4-phosphono-butyrate
700925
Methanocaldococcus jannaschii
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 33278, calculated from sequence; 1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Methanocaldococcus jannaschii
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
the heterologously expressed protein is thermostable
Methanocaldococcus jannaschii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at
Methanocaldococcus jannaschii
Other publictions for EC 6.3.2.32
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
700925
Li
Glutathione synthetase homolog ...
Methanocaldococcus jannaschii
Proc. Natl. Acad. Sci. USA
100
9785-9790
2003
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