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Literature summary 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

Activating Compound

EC Number Activating Compound Comment 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 Cloned (Comment) 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 Comment 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 Comment 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 Comment 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] Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment Textmining
6.3.2.32 Methanocaldococcus jannaschii Q58407
-
-
6.3.2.33 Methanocaldococcus jannaschii Q58037
-
-

Purification (Commentary)

EC Number Purification (Comment) 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] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 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 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 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 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 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 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 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 Methanocaldococcus jannaschii GDP + phosphate + 5,6,7,8-tetrahydrosarcinapterin
-
 ?

Subunits

EC Number Subunits Comment Organism
6.3.2.32 monomer 1 * 35000, SDS-PAGE Methanocaldococcus jannaschii
6.3.2.32 monomer 1 * 33278, calculated from sequence Methanocaldococcus jannaschii
6.3.2.33 homodimer 2 * 35000, SDS-PAGE Methanocaldococcus jannaschii
6.3.2.33 homodimer 2 * 33278, calculated from sequence Methanocaldococcus jannaschii

Synonyms

EC Number Synonyms Comment Organism
6.3.2.32 CofF
-
 Methanocaldococcus jannaschii
6.3.2.32 CofF protein
-
 Methanocaldococcus jannaschii
6.3.2.32 gamma-F420-2:alpha-L-glutamate ligase
-
 Methanocaldococcus jannaschii
6.3.2.32 MJ1001
-
 Methanocaldococcus jannaschii
6.3.2.33 H4MPT:alpha-L-glutamate ligase
-
 Methanocaldococcus jannaschii
6.3.2.33 MJ0620
-
 Methanocaldococcus jannaschii
6.3.2.33 MptN protein
-
 Methanocaldococcus jannaschii
6.3.2.33 tetrahydromethanopterin:alpha-L-glutamate ligase
-
 Methanocaldococcus jannaschii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment 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] Comment 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 Comment Organism
6.3.2.32 8.5
-
 assay at Methanocaldococcus jannaschii
6.3.2.33 7
-
 assay at Methanocaldococcus jannaschii