Information on EC 6.3.2.34 - coenzyme F420-1:gamma-L-glutamate ligase:
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The lowest common taxonomy group for this enzyme is: Euryarchaeota

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EC NUMBERCOMMENTARY
6.3.2.34-

RECOMMENDED NAMEGeneOntology No.
coenzyme F420-1:gamma-L-glutamate ligaseGO:0052619

REACTIONREACTION DIAGRAMCOMMENTARYORGANISM UNIPROT ACCESSION NO.LITERATURE
GTP + coenzyme F420-1 + L-glutamate = GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		----
GTP + coenzyme F420-1 + L-glutamate = GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		-Methanocaldococcus jannaschiiQ58178696202

REACTION TYPEORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

PATHWAYKEGG LinkMetaCyc Link
factor 420 polyglutamylation-PWY-5199

SYSTEMATIC NAMEIUBMB 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.

SYNONYMSORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
CofEMethanocaldococcus jannaschiiQ58178-696202
CofE-AFArchaeoglobus fulgidus DSM 4304O28028-699520
F420-0:gamma-glutamyl ligaseMethanocaldococcus jannaschiiQ58178-696202
F420:gamma-glutamyl ligaseArchaeoglobus fulgidus DSM 4304O28028family of non-ribosomal peptide synthases699520
MJ0768Methanocaldococcus jannaschiiQ58178-696202

CAS REGISTRY NUMBERCOMMENTARY
No entries in this field

ORGANISMCOMMENTARYLITERATURESEQUENCE CODESEQUENCE DB SOURCE
Archaeoglobus fulgidus DSM 4304-699520O28028SwissProtManually annotated by BRENDA team
Methanocaldococcus jannaschii-696202Q58178SwissProtManually annotated by BRENDA team

GENERAL INFORMATIONORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

SUBSTRATEPRODUCT                      REACTION DIAGRAMORGANISM UNIPROT ACCESSION NO. COMMENTARY/
Substrate		 LITERATURE/
Substrate		 COMMENTARY/
Product		 LITERATURE/
Product		 Reversibility
r=reversible
ir=irreversible
?=not specified
L-glutamate + dGTP + coenzyme F420-0UDP + phosphate + coenzyme gamma-F420-1
show the reaction diagram		Methanocaldococcus jannaschiiQ58178maximum 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 ligation696202--?
L-glutamate + dGTP + coenzyme gamma-F420-1dGDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Methanocaldococcus jannaschiiQ58178maximum 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 ligation696202--?
L-glutamate + GTP + coenzyme F420-1GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Archaeoglobus fulgidus DSM 4304O28028the 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 accumulated699520--?
L-glutamate + GTP + coenzyme gamma-F420-1GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Methanocaldococcus jannaschiiQ58178step in the biosynthesis of coenzyme F420, 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 ligation696202--?
L-glutamate + UTP + coenzyme gamma-F420-1UDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Methanocaldococcus jannaschiiQ58178maximum 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 ligation696202--?
additional information?-Methanocaldococcus jannaschiiQ58178CofE 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 residues696202---

NATURAL SUBSTRATESNATURAL PRODUCTSREACTION DIAGRAMORGANISM UNIPROT ACCESSION NO.COMMENTARY SUBSTRATELITERATURE
(Substrate)		COMMENTARY PRODUCTLITERATURE
(Product)
L-glutamate + GTP + coenzyme F420-1GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Archaeoglobus fulgidus DSM 4304O28028the 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 accumulated699520--
L-glutamate + GTP + coenzyme gamma-F420-1GDP + phosphate + coenzyme gamma-F420-2
show the reaction diagram		Methanocaldococcus jannaschiiQ58178step in the biosynthesis of coenzyme F420696202--

COFACTORORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATUREIMAGE
No entries in this field

METALS and IONS ORGANISM UNIPROT ACCESSION NO.COMMENTARY LITERATURE
Co2+Methanocaldococcus jannaschiiQ58178divalent 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 MnCl2696202
K+Methanocaldococcus jannaschiiQ58178CofE 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 effective696202
Mg2+Methanocaldococcus jannaschiiQ58178divalent 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 activity696202
Mn2+Methanocaldococcus jannaschiiQ58178divalent 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 activity696202
Mn2+Archaeoglobus fulgidus DSM 4304O28028there are two Mn2+-binding sites per monomer within close proximity of the GDP alpha and beta-phosphate groups699520
NH4+Methanocaldococcus jannaschiiQ58178CofE 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 effective696202

INHIBITORSORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
beta,gamma-CH2-GTPMethanocaldococcus jannaschiiQ581785 mM, 56% inhibition696202 2D-image
Ca2+Methanocaldococcus jannaschiiQ5817810 mM696202 2D-image
Cs2+Methanocaldococcus jannaschiiQ58178in the presence of either Rb+ or Cs+ at 0.2 M concentration no gamma-F420-2 is formed696202 2D-image
Ni2+Methanocaldococcus jannaschiiQ58178in the presence of NiCl2 no gamma-F420-2 is formed696202 2D-image
Rb2+Methanocaldococcus jannaschiiQ58178in the presence of either Rb+ or Cs+ at 0.2 M concentration no gamma-F420-2 is formed696202-
Zn2+Methanocaldococcus jannaschiiQ5817810 mM696202 2D-image
GDPMethanocaldococcus jannaschiiQ581785 mM, 67% inhibition696202 2D-image
additional informationMethanocaldococcus jannaschiiQ58178no 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 acid696202-

ACTIVATING COMPOUNDORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
additional informationMethanocaldococcus jannaschiiQ58178no change of CofE activity is observed when the enzyme is assayed with the addition of 10 mM dithiothreitol to the reaction mixture696202-

KM VALUE [mM]KM VALUE [mM] MaximumSUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
0.00021-coenzyme gamma-F420-1Methanocaldococcus jannaschiiQ58178pH 8.5, 50°C696202 2D-image

TURNOVER NUMBER [1/s] TURNOVER NUMBER MAXIMUM[1/s] SUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

kcat/KM VALUE [1/mMs-1]kcat/KM VALUE [1/mMs-1] MaximumSUBSTRATEORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

Ki VALUE [mM]Ki VALUE [mM] MaximumINHIBITORORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

IC50 VALUE [mM]IC50 VALUE [mM] MaximumINHIBITORORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE IMAGE
No entries in this field

SPECIFIC ACTIVITY [µmol/min/mg] SPECIFIC ACTIVITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

pH OPTIMUMpH MAXIMUMORGANISM UNIPROT ACCESSION NO. COMMENTARYLITERATURE
8.5-Methanocaldococcus jannaschiiQ58178with 50 mM CHES/Na+ buffer696202
8.5-Archaeoglobus fulgidus DSM 4304O28028assay at699520

pH RANGEpH RANGE MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

TEMPERATURE OPTIMUMTEMPERATURE OPTIMUM MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
50-Archaeoglobus fulgidus DSM 4304O28028assay at699520
60-Methanocaldococcus jannaschiiQ58178-696202

TEMPERATURE RANGE TEMPERATURE MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
3580Methanocaldococcus jannaschiiQ5817835°C: about 30% of maximal activity, 80°C: about 55% of maximal activity696202

pI VALUEpI VALUE MAXIMUMORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

SOURCE TISSUE ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE SOURCE
No entries in this field

LOCALIZATION ORGANISM UNIPROT ACCESSION NO. COMMENTARY GeneOntology No. LITERATURE SOURCE
No entries in this field

PDBSCOPCATHORGANISM
No entries in this field

MOLECULAR WEIGHT MOLECULAR WEIGHT MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
52100-Methanocaldococcus jannaschiiQ58178gel filtration696202

SUBUNITS ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
homodimerMethanocaldococcus jannaschiiQ581782 * 27150, calculated from sequence696202

POSTTRANSLATIONAL MODIFICATION ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

Crystallization/COMMENTARY ORGANISM UNIPROT ACCESSION NO. 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°CArchaeoglobus fulgidus DSM 4304O28028699520

pH STABILITYpH STABILITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

TEMPERATURE STABILITYTEMPERATURE STABILITY MAXIMUM ORGANISM UNIPROT ACCESSION NO. COMMENTARYLITERATURE
80-Methanocaldococcus jannaschiiQ5817815 min, 30% loss of activity696202

GENERAL STABILITYORGANISM UNIPROT ACCESSION NO.LITERATURE
No entries in this field

ORGANIC SOLVENT ORGANISM UNIPROT ACCESSION NO. COMMENTARY LITERATURE
No entries in this field

OXIDATION STABILITY ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

STORAGE STABILITY ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

Purification/COMMENTARY ORGANISM UNIPROT ACCESSION NO. LITERATURE
-Archaeoglobus fulgidus DSM 4304O28028699520
recombinant enzymeMethanocaldococcus jannaschiiQ58178696202

Cloned/COMMENTARY ORGANISM UNIPROT ACCESSION NO. LITERATURE
-Archaeoglobus fulgidus DSM 4304O28028699520
expressed in Escherichia coliMethanocaldococcus jannaschiiQ58178696202

EXPRESSION ORGANISM UNIPROT ACCESSION NO. LITERATURE
No entries in this field

ENGINEERINGORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

Renatured/COMMENTARYORGANISM UNIPROT ACCESSION NO.LITERATURE
No entries in this field

APPLICATIONORGANISM UNIPROT ACCESSION NO.COMMENTARYLITERATURE
No entries in this field

REF. AUTHORS TITLE JOURNAL VOL. PAGES YEAR ORGANISMLINK TO PUBMEDSOURCE
696202Li, H.; Graupner, M.; Xu, H.; White, R.H.CofE catalyzes the addition of two glutamates to F420-0 in F420 coenzyme biosynthesis in Methanococcus jannaschiiBiochemistry429771-97782003Methanocaldococcus jannaschii PubMed
699520Nocek, 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 synthasesJ. Mol. Biol.372456-4692007Archaeoglobus fulgidus DSM 4304 PubMed

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KEGG
MetaCyc
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
SYSTERS
Protein Mutant Database
InterPro (database of protein families, domains and functional sites)