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Literature summary for 6.3.2.32 extracted from

  • Li, H.; Xu, H.; Graham, D.E.; White, R.H.
    Glutathione synthetase homologs encode alpha-L-glutamate ligases for methanogenic coenzyme F420 and tetrahydrosarcinapterin biosyntheses (2003), Proc. Natl. Acad. Sci. USA, 100, 9785-9790.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.2.32

Activating Compound

Activating Compound Comment 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 (Comment) Organism
expression in Escherichia coli Methanocaldococcus jannaschii

Inhibitors

Inhibitors Comment 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 Comment Organism Structure
0.0012
-
 coenzyme gamma-F420-2 pH 8.5, 50°C Methanocaldococcus jannaschii

Metals/Ions

Metals/Ions Comment 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] Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 Comment Textmining
Methanocaldococcus jannaschii Q58407
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 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 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 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 Methanocaldococcus jannaschii ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 35000, SDS-PAGE Methanocaldococcus jannaschii
monomer 1 * 33278, calculated from sequence Methanocaldococcus jannaschii

Synonyms

Synonyms Comment 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] Comment Organism
50
-
 assay at Methanocaldococcus jannaschii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 the heterologously expressed protein is thermostable Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Methanocaldococcus jannaschii