Information on EC 4.1.99.22 - GTP 3',8-cyclase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.1.99.22
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RECOMMENDED NAME
GeneOntology No.
GTP 3',8-cyclase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GTP + S-adenosyl-L-methionine + reduced electron acceptor = (8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate + 5'-deoxyadenosine + L-methionine + oxidized electron acceptor
show the reaction diagram
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GTP = cyclic pyranopterin phosphate + diphosphate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Folate biosynthesis
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Metabolic pathways
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molybdenum cofactor biosynthesis
SYSTEMATIC NAME
IUBMB Comments
GTP 3',8-cyclase [(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate-forming]
The enzyme catalyses an early step in the biosynthesis of the molybdenum cofactor (MoCo). In bacteria and plants the reaction is catalysed by MoaA and Cnx2, respectively. In mammals it is catalysed by the MOCS1A domain of the bifunctional MOCS1 protein, which also catalyses EC 4.6.1.17, cyclic pyranopterin monophosphate synthase. The enzyme belongs to the superfamily of radical S-adenosyl-L-methionine (radical SAM) enzymes, and contains two oxygen-sensitive FeS clusters.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
mog gene, aq_061
UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
gene TTHA0341
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate
cyclic pyranopterin phosphate + diphosphate
show the reaction diagram
GMP[CH2]PP + S-adenosyl-L-methionine + reduced electron acceptor
(8S)-3',8-cyclo-7,8-dihydroguanosine-P[CH2]PP + 5'-deoxyadenosine + L-methionine + oxidized electron acceptor
show the reaction diagram
GTP
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate
show the reaction diagram
GTP
cyclic pyranopterin monophosphate + diphosphate
show the reaction diagram
GTP
cyclic pyranopterin phosphate + diphosphate
show the reaction diagram
GTP + S-adenosyl-L-methionine + reduced electron acceptor
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate + 5'-deoxyadenosine + L-methionine + oxidized electron acceptor
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate
cyclic pyranopterin phosphate + diphosphate
show the reaction diagram
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reaction of MoaC
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GTP
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate
show the reaction diagram
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reaction of MoaA with GTP, S-adenosyl-L-methionine, and sodium dithionite in the absence of MoaC
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?
GTP
cyclic pyranopterin monophosphate + diphosphate
show the reaction diagram
GTP
cyclic pyranopterin phosphate + diphosphate
show the reaction diagram
GTP + S-adenosyl-L-methionine + reduced electron acceptor
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate + 5'-deoxyadenosine + L-methionine + oxidized electron acceptor
show the reaction diagram
additional information
?
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MoaA catalyzes a unique radical C-C bond formation reaction via a 5'-deoxyadenosyl radical intermediate and that, in contrast to previous proposals, MoaC plays a major role in the complex rearrangement to generate the pyranopterin ring
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
iron-sulfur centre
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
iron-sulfur centre
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
MoaC protein
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00006 - 0.00079
(8S)-3',8-cyclo-7,8-dihydroguanosine 5'-triphosphate
0.0031 - 0.0069
GTP
0.0051 - 0.036
S-adenosyl-L-methionine
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00008 - 0.0007
GTP
0.00007 - 0.00075
S-adenosyl-L-methionine
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41000
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monomer, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da), gel filtration
42000
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1 * 42000, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da); 2 * 42000, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da)
82000
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homodimer, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da), gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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2 * 42000, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da)
monomer
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1 * 42000, in solution MoaA exists as a monomer (41000 Da) and dimer (82000 Da)
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme, mixing of 0.001 ml of 24 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 0.2 M NaCl, with 0.001 ml of reservoir solution containing 40% v/v PEG 600, 100 mM CHES buffer, pH 9.5, or 0.2 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, 25% w/v PEG 3350, 1 week, two different crystal forms, X-ray diffraction structure determination and analysis at 1.7-1.9 A resolution, molecular replacement method
crystal structure of wild-type MoaA, MoaA-R17A/R266A/R268A and MoaA in complex with 5'-GTP2.35 A resolution
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crystals are grown under anaerobic conditions, hanging drop vapor diffusion technique, crystals belong to space group P2(1)2(1)2(1) with cell dimensions of a = 48.1, b = 102.4, and c = 191.2 A and contain two molecules per asymmetric unit, structures of MoaA in the apo-state (2.8 A) and in complex with S-adenosyl-L-methionine (2.2 A)
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purified recombinant enzyme, X-ray diffraction structure determination and analysis at 1.64 A resolution, molecular replacement method
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified under anaerobic conditions
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recombinant enzyme
recombinant enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIL by two different times of gel filtration and anion exchange chromatography, followed by gel filtration and hydroxyapatite chromatography, and a last step of gel filtration
recombinant MOCS1B from Escherichia coli strain BL21(DE3) by streptomycin sulfate and ammonium sulfate precipitation steps, nickel affinity chromatography, gell filtration, and ultrafiltration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as His-tagged proteins in Escherichia coli
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expression in Escherichia coli
gene mog, phylogenetic tree, expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3)-RIL
gene mogA, phylogenetic tree, recombinant expression
MOCS1B overexpression in Escherichia coli strain BL21(DE3)
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the first two enzymes of the molybdopterin biosynthesis pathway are encoded by the MOCS1 locus in humans. Isoform MOCS1A and isoform MOCS1B form a complex that catalyzes the conversion of 5'-GTP to cyclic pyranopterin monophosphate. They are encoded by an apparently bicistronic MOCS1AMOCS1B transcript which bypasses the normal termination nonsense codon of MOCS1A resulting in fusion of the MOCS1A and MOCS1B open reading frames. The bicistronic form of MOCS1 mRNA is likely to only produce MOCS1A protein and suggests that MOCS1B is translated only as a fusion with MOCS1A
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C24S/C28S/C31S
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the mutant does not contain the catalytic S-adenosyl-L-methionine-binding cluster I
D198A
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mutation in GG motif, decrease in affinity for S-adenosine-L-methionine
G339A
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mutation in GG motif, loss of activity
G340A
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mutation in GG motif, loss of activity
N124A/N165A
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mutation reduces binding of 5'-GTP
R17A
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complete loss of activity
R17A/R266A/R268A
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complete loss of activity
R192A
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80% loss of activity
R266A
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complete loss of activity
R268A
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complete loss of activity
R71A
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80% loss of activity
S126A
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mutant enzyme with low activity
T73A
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mutant enzyme with low activity
Y30A
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mutant enzyme with low activity
D198A
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mutation in GG motif, decrease in affinity for S-adenosine-L-methionine
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G339A
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mutation in GG motif, loss of activity
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G340A
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mutation in GG motif, loss of activity
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