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Literature summary extracted from

  • Graham, D.E.; Xu, H.; White, R.H.
    A member of a new class of GTP cyclohydrolases produces formylaminopyrimidine nucleotide monophosphates (2002), Biochemistry, 41, 15074-15084.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.4.29 NH4+ activation of GTP cyclohydrolase activity, maximal activity at 50 mM Methanocaldococcus jannaschii

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.4.29 expression in Escherichia coli Methanocaldococcus jannaschii

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.29 H136Q approx. 7 and 15% of wild-type GTP cyclohydrolase and diphosphate phosphohydrolase activity, respectively Methanocaldococcus jannaschii

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.4.29 8-azido-GTP 0.6 mM, 91% inhibition of GTP cyclohydrolase activity, 0.3 mM, 53% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 8-bromo-GTP 0.1 mM, 50% inhibition of GTP cyclohydrolase activity, 0.1 mM, 64% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 alpha,beta-methylene-GTP 0.06 mM, 80% inhibition of GTP cyclohydrolase activity, 0.6 mM, 33% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 beta,gamma-methylene-GTP 0.6 mM, 34% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 diphosphate inhibition of diphosphate phosphohydrolase activity at high concentrations Methanocaldococcus jannaschii
3.5.4.29 GDP more than 40% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 GMP more than 40% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 GTP-gamma-S 0.6 mM, 80% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 IMP 0.6 mM, 25% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 methylene diphosphate 0.6 mM, 40% inhibition of GTP cyclohydrolase activity, 0.6 mM, 56% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 NaF 0.06 mM, 55% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 phosphate 0.6 mM, 25% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 tripolyphosphate 0.6 mM, 50% inhibition of GTP cyclohydrolase activity, 0.05 mM, 52% inhibition of diphosphate phosphohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 XTP 0.6 mM, 52% inhibition of GTP cyclohydrolase activity Methanocaldococcus jannaschii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.29 0.054
-
 GTP pH 8.0, 70°C, GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 0.063
-
 GTP pH 8.0, 70°C, diphosphate phosphohydrolase activity Methanocaldococcus jannaschii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.4.29 CoCl2 45% of Mg2+ diphosphate phosphohydrolase activation Methanocaldococcus jannaschii
3.5.4.29 K+ activation of GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 Mg2+ required for cyclohydrolase activity, maximal activity at 4 mM, no other metal supports full activity, maximal diphosphate phosphohydrolase activity in the presence of 5 mM MgCl2 Methanocaldococcus jannaschii
3.5.4.29 Mn2+ 7% of Mg2+ activation, 8% of Mg2+ diphosphate phosphohydrolase activation Methanocaldococcus jannaschii
3.5.4.29 Zn(OAc)2 21% of Mg2+ diphosphate phosphohydrolase activation Methanocaldococcus jannaschii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.4.29 30275
-
 3 * 30275, deduced from nucleotide sequence Methanocaldococcus jannaschii
3.5.4.29 30359
-
 3 * 30359, MALDI-TOF mass spectrometry Methanocaldococcus jannaschii
3.5.4.29 37000
-
 3 * 37000, SDS-PAGE Methanocaldococcus jannaschii
3.5.4.29 98300
-
 gel filtration Methanocaldococcus jannaschii

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.29 Methanocaldococcus jannaschii Q57609 hyperthermophilic euryarcheon
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.29 heating, streptomycin, Mono Q, Sephacryl, recombinant enzyme Methanocaldococcus jannaschii

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.4.29 GTP + 3 H2O = 2-amino-5-formylamino-6-(5-phospho-D-ribosylamino)pyrimidin-4(3H)-one + 2 phosphate although phosphate is the product of the enzymes phosphohydrolase activity, diphosphate is a likely intermediate Methanocaldococcus jannaschii
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.4.29 0.39
-
 recombinant enzyme Methanocaldococcus jannaschii

Storage Stability

EC Number Storage Stability Organism
3.5.4.29 4°C, 50 mM TAPS/KOH, pH 8.0, 150 mM KCl, 9 weeks, 20% loss of activity Methanocaldococcus jannaschii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.29 beta,gamma-methylene-GTP + H2O
-
 Methanocaldococcus jannaschii 2-amino-5-formylamino-6-hydroxy-4-(5-phosphoribosylamino)-pyrimidine + methyl phosphonate
-
 ?
3.5.4.29 dGTP + H2O
-
 Methanocaldococcus jannaschii 2-amino-5-formylamino-6-hydroxy-4-(deoxy-5-phosphoribosylamino)-pyrimidine + phosphate
-
 ?
3.5.4.29 diphosphate + H2O
-
 Methanocaldococcus jannaschii 2 phosphate
-
 ?
3.5.4.29 GTP + H2O
-
 Methanocaldococcus jannaschii 2-amino-5-formylamino-6-hydroxy-4-(5-phosphoribosylamino)-pyrimidine + phosphate enzyme has diphosphate phosphohydrolase and GTP cyclohydrolase activities, no activity with 8-bromo-GTP, 8-azido-GTP, alpha,beta-methylene-GTP, ATP, GDP, GMP and XTP ?
3.5.4.29 GTP-gamma-S + H2O
-
 Methanocaldococcus jannaschii 2-amino-5-formylamino-6-hydroxy-4-(5-phosphoribosylamino)-pyrimidine + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.4.29 trimer 3 * 37000, SDS-PAGE Methanocaldococcus jannaschii
3.5.4.29 trimer 3 * 30275, deduced from nucleotide sequence Methanocaldococcus jannaschii
3.5.4.29 trimer 3 * 30359, MALDI-TOF mass spectrometry Methanocaldococcus jannaschii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.4.29 70
-
 GTP hydrolase and diphosphate phosphohydrolase activity Methanocaldococcus jannaschii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.4.29 0.35
-
 GTP pH 8.0, 70°C, GTP cyclohydrolase activity Methanocaldococcus jannaschii
3.5.4.29 0.5
-
 GTP pH 8.0, 70°C, diphosphate phosphohydrolase activity Methanocaldococcus jannaschii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.4.29 8 9 GTP hydrolase and diphosphate phosphohydrolase activity Methanocaldococcus jannaschii

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.5.4.29 0.187
-
 diphosphate
-
 Methanocaldococcus jannaschii