This protein catalyses the successive addition of two glutamate residues to cofactor F420 by two distinct and independent reactions. In the first reaction (EC 6.3.2.31, coenzyme F420-0---L-glutamate ligase) the enzyme attaches a glutamate via its alpha-amine group to F420-0. In the second reaction, which is described here, the enzyme catalyses the addition of a second L-glutamate residue to the gamma-carboxyl of the first glutamate.
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The expected taxonomic range for this enzyme is: Bacteria, Archaea
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SYSTEMATIC NAME
IUBMB Comments
L-glutamate:coenzyme F420-1 ligase (GDP-forming)
This protein catalyses the successive addition of two glutamate residues to cofactor F420 by two distinct and independent reactions. In the first reaction (EC 6.3.2.31, coenzyme F420-0---L-glutamate ligase) the enzyme attaches a glutamate via its alpha-amine group to F420-0. In the second reaction, which is described here, the enzyme catalyses the addition of a second L-glutamate residue to the gamma-carboxyl of the first glutamate.
maximum activity for coenzyme F420-2 formation is 25% of the maximum activity with GTP. Coenzyme F420-1 is coenzyme F420 with one glutamic acid residue (attached via its alpha-NH2 to F420-0), the second glutamate in bound via a gamma ligation
maximum activity for coenzyme F420-2 formation is 25% of the maximum activity with GTP. Coenzyme F420-1 is coenzyme F420 with one glutamic acid residue (attached via its alpha-NH2 to F420-0), the second glutamate in bound via a gamma ligation
the enzyme protein catalyzes two distinct and independent reactions, firstly attaching a glutamte via its alpha-NH2 to F420-0 (cf. coenzyme F420-0:glutamyl ligase). The second reaction is a gamma ligation, taking place when a certain amount of monoglutamylated F420-1 has accumulated
maximum activity for coenzyme F420-2 formation is 66% of the maximum activity with GTP. Coenzyme F420-1 is coenzyme F420 with one glutamic acid residue (attached via its alpha-NH2 to F420-0), the second glutamate in bound via a gamma ligation
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
coenzyme F420-1 is coenzyme F420 with one glutamic acid residue (attached via its alpha-NH2 to F420-0), the second glutamate in bound via a gamma ligation
CofE incubated with 10 mM beta-glutamate, D-glutamate, gamma-glutamylglutamate, DL-2-amino-3-phosphonopropionic acid, 2-carboxyethylphosphonic acid, or L-R-aminoadipic acid produces no F420-2 or other F420 analogues. CofE cannot use F420-2 as substrate to add more glutamate residues
full-length FbiB adds multiple L-glutamate residues to F420-0 in vitro to produce F420-5, i.e. 5 L-glutamate residues in the poly-gamma-glutamate tail after 24 h, reactions of EC 6.3.2.31 and 6.3.2.34. Communication between the two domains of the protein is critical for full gamma-glutamyl ligase activity. No observed activity for dGTP, ATP, and dATP nucleotides
full-length FbiB adds multiple L-glutamate residues to F420-0 in vitro to produce F420-5, i.e. 5 L-glutamate residues in the poly-gamma-glutamate tail after 24 h, reactions of EC 6.3.2.31 and 6.3.2.34. Communication between the two domains of the protein is critical for full gamma-glutamyl ligase activity. No observed activity for dGTP, ATP, and dATP nucleotides
full-length FbiB adds multiple L-glutamate residues to F420-0 in vitro to produce F420-5, i.e. 5 L-glutamate residues in the poly-gamma-glutamate tail after 24 h, reactions of EC 6.3.2.31 and 6.3.2.34. Communication between the two domains of the protein is critical for full gamma-glutamyl ligase activity. No observed activity for dGTP, ATP, and dATP nucleotides
the enzyme protein catalyzes two distinct and independent reactions, firstly attaching a glutamte via its alpha-NH2 to F420-0 (cf. coenzyme F420-0:glutamyl ligase). The second reaction is a gamma ligation, taking place when a certain amount of monoglutamylated F420-1 has accumulated
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
reduced coenzyme F420 (F420H2) is the true substrate of F420-0-gamma-glutamyl ligase for the attachment of more than two glutamate residues to the coenzyme
divalent cation requirement, maximum CofE activity is observed with the addition of 10 mM MnCl2. Reactions containing 10 mM either MgCl2 or CoCl2 produce 40% of the F420-2 of that is produced with MnCl2
CofE absolutely requires a monovalent cation for activity, the greatest extent of activation is achieved by K+, with maximum stimulation occurring at 0.2 M KCl, NH4+ stimulates activity to a lesser extent, extent, whereas Na+ and Li+ have no effect on CofE activity. A mixture of Mn2+, Mg2+, and K+ is the most effective
divalent cation requirement, maximum CofE activity is observed with the addition of 10 mM MnCl2. Reactions containing 10 mM either MgCl2 or CoCl2 produce 40% of the F420-2 of that is produced with MnCl2. The combination of 5 mM MgCl2 and 2-5 mM of MnCl2 supports the highest activity
CofE absolutely requires a monovalent cation for activity, the greatest extent of activation is achieved by K+, NH4+ stimulates activity to a lesser extent, whereas Na+ and Li+ have no effect on CofE activity. A mixture of Mn2+, Mg2+, and K+ is the most effective
divalent cation requirement, maximum CofE activity is observed with the addition of 10 mM MnCl2. Reactions containing 10 mM either MgCl2 or CoCl2 produce 40% of the F420-2 of that is produced with MnCl2. The combination of 5 mM MgCl2 and 2-5 mM of MnCl2 supports the highest activity
no significant inhibition when CofE is incubated with the following compound (10 mM): L-aspartate, L-glutamine, L-homocysteic acid, or DL-amino-4-phosphono-butyric acid
both genes fbiA and fbiB are essential for normal coenzyme F420-5,6 production. fbiA and fbiB constitute an operon. Very low levels of fbiB mRNA are produced by the fbiA mutant
FbiB produces cofactor F420 with predominantly 5-7 L-glutamate residues in the poly-gamma-glutamate tail, reactions of EC 6.3.2.31 and 6.3.2.34. The N-terminal domain of FbiB is homologous to CofE with an annotated gamma-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Communication between the two domains is critical for full gamma-glutamyl ligase activity
in vitro, MtbFbiB synthesizes side chains containing up to seven glutamate residues if cofactor F420 is presented to the enzyme in a two-electron reduced state (F420H2). The polyglutamylation process requires the assistance of F420-dependent glucose-6-phosphate dehydrogenase (Fgd) which reduces F420 to F420H2. Starting with F420-0H2, the amino-terminal domain of FbiB may build F420-2H2, which is then transferred to the carboxy-terminal domain for further glutamylation. F420-2H2 modifies the carboxy-terminal domain structurally to accommodate longer glutamyl chains
both genes fbiA and fbiB are essential for normal coenzyme F420-5,6 production. fbiA and fbiB constitute an operon. Very low levels of fbiB mRNA are produced by the fbiA mutant
FbiB produces cofactor F420 with predominantly 5-7 L-glutamate residues in the poly-gamma-glutamate tail, reactions of EC 6.3.2.31 and 6.3.2.34. The N-terminal domain of FbiB is homologous to CofE with an annotated gamma-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Communication between the two domains is critical for full gamma-glutamyl ligase activity
FbiB produces cofactor F420 with predominantly 5-7 L-glutamate residues in the poly-gamma-glutamate tail, reactions of EC 6.3.2.31 and 6.3.2.34. The N-terminal domain of FbiB is homologous to CofE with an annotated gamma-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Communication between the two domains is critical for full gamma-glutamyl ligase activity
in vitro, MtbFbiB synthesizes side chains containing up to seven glutamate residues if cofactor F420 is presented to the enzyme in a two-electron reduced state (F420H2). The polyglutamylation process requires the assistance of F420-dependent glucose-6-phosphate dehydrogenase (Fgd) which reduces F420 to F420H2. Starting with F420-0H2, the amino-terminal domain of FbiB may build F420-2H2, which is then transferred to the carboxy-terminal domain for further glutamylation. F420-2H2 modifies the carboxy-terminal domain structurally to accommodate longer glutamyl chains
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of the enzyme from Archaeoglobus fulgidus and its complex with GDP at 2.5 A and 1.35 A resolution, respectively. CofE-AF crystallization is performed by the sitting-drop and hanging-drop methods using vapor diffusion at 18°C
structures of the C-terminal domain of FbiB in apo-, F420-0-, and FMN-bound states, displaying distinct sites for F420-0 and FMN ligands that partially overlap
Nocek, B.; Evdokimova, E.; Proudfoot, M.; Kudritska, M.; Grochowski, L.L.; White, R.H.; Savchenko, A.; Yakunin, A.F.; Edwards, A.; Joachimiak, A.
Structure of an amide bond forming F(420):gamma-glutamyl ligase from Archaeoglobus fulgidus - a member of a new family of non-ribosomal peptide synthases
Use of transposon Tn5367 mutagenesis and a nitroimidazopyran-based selection system to demonstrate a requirement for fbiA and fbiB in coenzyme F420 biosynthesis by Mycobacterium bovis BCG