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Literature summary extracted from

  • Butterworth, A.C.; Medrano, F.J.; Eakin, A.E.; Craig, S.P., 3rd
    Saturation mutagenesis, complement selection, and steady-state kinetic studies illuminate the roles of invariant residues in active site loop I of the hypoxanthine phosphoribosyltransferase from Trypanosoma cruzi (2004), Biochim. Biophys. Acta, 1699, 87-94.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.4.2.8 pharmacology enzyme is a potential drug target in the treatment of parasite caused disease Trypanosoma cruzi

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.8 expression of wild-type and mutant enzymes in Escherichia coli Trypanosoma cruzi

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.8 G69S reduced activity compared to the wild-type enzyme Trypanosoma cruzi
2.4.2.8 L67M highly reduced activity with hypoxanthine compared to the wild-type enzyme, mutant is not active with guanine Trypanosoma cruzi
2.4.2.8 additional information saturation mutagenesis and complement selection for investigation of functional roles of residues Leu67 and Gly69, sequencing of 70 clones and identification of 30 different mutations, several mutants of L67 or G69 support bacterial growth on minimal medium, but only L67M and G69S can be expressed and purified from overexpressing bacteria, overview Trypanosoma cruzi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.8 0.0086
-
 hypoxanthine pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 0.012
-
 guanine pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 0.0192
-
 hypoxanthine pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.022
-
 guanine pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.027
-
 IMP pH 7.5, 37°C, recombinant wild-type enzyme and mutant L67M Trypanosoma cruzi
2.4.2.8 0.032
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 0.0339
-
 hypoxanthine pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.035
-
 diphosphate pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 0.08
-
 IMP pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.083
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.132
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.135
-
 diphosphate pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.214
-
 diphosphate pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.8 Mg2+ at 12 mM Trypanosoma cruzi

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.8 GMP + diphosphate Trypanosoma cruzi
-
 guanine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 r
2.4.2.8 IMP + diphosphate Trypanosoma cruzi
-
 hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.8 Trypanosoma cruzi
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.8 recombinant wild-type and mutant enzymes from Escherichia coli by GMP affinity and cation exchange chromatography Trypanosoma cruzi

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.2.8 IMP + diphosphate = hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate reaction mechanism, function and influence of invariant active site loop I residues on enzyme efficiency in forward and reverse reaction Trypanosoma cruzi
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.8 additional information
-
-
 Trypanosoma cruzi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.8 GMP + diphosphate
-
 Trypanosoma cruzi guanine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 r
2.4.2.8 IMP + diphosphate
-
 Trypanosoma cruzi hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 r
2.4.2.8 additional information L67 side chain participates in hydrophobic interactions between dimer subunits important for the proper positioning of main chain atoms that influence enzyme chemistry and the binding of 5-phospho-alpha-D-ribose 1-diphosphate, diphosphate, and hypoxanthine, residue 69 needs to be small like Gly, which is found there, to avoid sterical interference with binding of 5-phospho-alpha-D-ribose 1-diphosphate and diphosphate as well as the positioning of a metal ion Trypanosoma cruzi ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.2.8 HPRT
-
 Trypanosoma cruzi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.8 37
-
 assay at Trypanosoma cruzi

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.8 0.07
-
 IMP pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.12
-
 diphosphate pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.19
-
 IMP pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.3
-
 hypoxanthine pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 0.31
-
 diphosphate pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 0.47
-
 IMP pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 0.48
-
 diphosphate pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 4.7
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant mutant L67M Trypanosoma cruzi
2.4.2.8 20
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 22
-
 hypoxanthine pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 22.1
-
 guanine pH 7.5, 37°C, recombinant mutant G69S Trypanosoma cruzi
2.4.2.8 22.9
-
 hypoxanthine pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 23.2
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi
2.4.2.8 32.9
-
 guanine pH 7.5, 37°C, recombinant wild-type enzyme Trypanosoma cruzi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.8 7.5
-
 assay at Trypanosoma cruzi