BRENDA - Enzyme Database

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
EC Number
Activating Compound
Commentary
Organism
Structure
6.3.2.32
2-mercaptoethanol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
6.3.2.32
dithiothreitol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
Cloned(Commentary)
EC Number
Commentary
Organism
6.3.2.32
expression in Escherichia coli
Methanocaldococcus jannaschii
6.3.2.33
expression in Escherichia coli
Methanocaldococcus jannaschii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
6.3.2.32
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
6.3.2.32
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
6.3.2.32
additional information
no inhibition by beta,gamma-CH2-ATP
Methanocaldococcus jannaschii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.2.32
0.0012
-
coenzyme gamma-F420-2
pH 8.5, 50°C
Methanocaldococcus jannaschii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.2.32
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
6.3.2.32
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
6.3.2.33
Mg2+
conserved Mg-ATP-binding domain
Methanocaldococcus jannaschii
6.3.2.33
additional information
does not require K+ for activity
Methanocaldococcus jannaschii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.3.2.32
33278
-
1 * 33278, calculated from sequence
Methanocaldococcus jannaschii
6.3.2.32
35000
-
1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.32
37000
-
gel filtration
Methanocaldococcus jannaschii
6.3.2.33
33278
-
2 * 33278, calculated from sequence
Methanocaldococcus jannaschii
6.3.2.33
35000
-
2 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.33
72000
-
gel filtration
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.2.32
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
-
-
?
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
Methanocaldococcus jannaschii
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
Methanocaldococcus jannaschii
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.3.2.32
Methanocaldococcus jannaschii
Q58407
-
-
6.3.2.33
Methanocaldococcus jannaschii
Q58037
-
-
Purification (Commentary)
EC Number
Commentary
Organism
6.3.2.32
recombinant enzyme
Methanocaldococcus jannaschii
6.3.2.33
-
Methanocaldococcus jannaschii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
6.3.2.33
12
-
under rate-limiting assay conditions, MptN produces 12.7 nmol of sarcinapterin per min per mg of protein in the presence of ATP and K+
Methanocaldococcus jannaschii
6.3.2.33
12.7
-
under rate-limiting assay conditions, MptN produces 12.7 nmol of sarcinapterin per min per mg of protein in the presence of GTP and K+
Methanocaldococcus jannaschii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.2.32
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
-
-
-
?
6.3.2.32
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
-
-
-
?
6.3.2.32
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
-
-
-
?
6.3.2.32
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
?
-
-
-
-
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
6.3.2.32
monomer
1 * 33278, calculated from sequence; 1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.33
homodimer
2 * 33278, calculated from sequence; 2 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.3.2.32
50
-
assay at
Methanocaldococcus jannaschii
6.3.2.33
60
-
assay at
Methanocaldococcus jannaschii
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
6.3.2.32
additional information
-
the heterologously expressed protein is thermostable
Methanocaldococcus jannaschii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.2.32
8.5
-
assay at
Methanocaldococcus jannaschii
6.3.2.33
7
-
assay at
Methanocaldococcus jannaschii
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
6.3.2.32
2-mercaptoethanol
without 2 mM dithiothreirol or 2-mercaptoethanol in the reaction mixture, CofF activity is up to 5fold lower
Methanocaldococcus jannaschii
6.3.2.32
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)
EC Number
Commentary
Organism
6.3.2.32
expression in Escherichia coli
Methanocaldococcus jannaschii
6.3.2.33
expression in Escherichia coli
Methanocaldococcus jannaschii
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
6.3.2.32
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
6.3.2.32
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
6.3.2.32
additional information
no inhibition by beta,gamma-CH2-ATP
Methanocaldococcus jannaschii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.2.32
0.0012
-
coenzyme gamma-F420-2
pH 8.5, 50°C
Methanocaldococcus jannaschii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.2.32
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
6.3.2.32
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
6.3.2.33
Mg2+
conserved Mg-ATP-binding domain
Methanocaldococcus jannaschii
6.3.2.33
additional information
does not require K+ for activity
Methanocaldococcus jannaschii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.3.2.32
33278
-
1 * 33278, calculated from sequence
Methanocaldococcus jannaschii
6.3.2.32
35000
-
1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.32
37000
-
gel filtration
Methanocaldococcus jannaschii
6.3.2.33
33278
-
2 * 33278, calculated from sequence
Methanocaldococcus jannaschii
6.3.2.33
35000
-
2 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.33
72000
-
gel filtration
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.2.32
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
-
-
?
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
Methanocaldococcus jannaschii
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
Methanocaldococcus jannaschii
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.2.32
recombinant enzyme
Methanocaldococcus jannaschii
6.3.2.33
-
Methanocaldococcus jannaschii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
6.3.2.33
12
-
under rate-limiting assay conditions, MptN produces 12.7 nmol of sarcinapterin per min per mg of protein in the presence of ATP and K+
Methanocaldococcus jannaschii
6.3.2.33
12.7
-
under rate-limiting assay conditions, MptN produces 12.7 nmol of sarcinapterin per min per mg of protein in the presence of GTP and K+
Methanocaldococcus jannaschii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.2.32
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
-
-
-
?
6.3.2.32
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
-
-
-
?
6.3.2.32
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
-
-
-
?
6.3.2.32
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
?
-
-
-
-
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
ATP + tetrahydromethanopterin + L-glutamate
the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
ADP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
biosynthesis of the cofactor 5,6,7,8-tetrahydrosarcinapterin, the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
6.3.2.33
GTP + tetrahydromethanopterin + L-glutamate
the enzyme does not discriminate between ATP and GTP
700925
Methanocaldococcus jannaschii
GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
6.3.2.32
monomer
1 * 33278, calculated from sequence; 1 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
6.3.2.33
homodimer
2 * 33278, calculated from sequence; 2 * 35000, SDS-PAGE
Methanocaldococcus jannaschii
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.3.2.32
50
-
assay at
Methanocaldococcus jannaschii
6.3.2.33
60
-
assay at
Methanocaldococcus jannaschii
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
6.3.2.32
additional information
-
the heterologously expressed protein is thermostable
Methanocaldococcus jannaschii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.2.32
8.5
-
assay at
Methanocaldococcus jannaschii
6.3.2.33
7
-
assay at
Methanocaldococcus jannaschii