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Literature summary extracted from

  • Valsecchi, W.M.; Cousido-Siah, A.; Defelipe, L.A.; Mitschler, A.; Podjarny, A.; Santos, J.; Delfino, J.M.
    The role of the C-terminal region on the oligomeric state and enzymatic activity of Trypanosoma cruzi hypoxanthine phosphoribosyl transferase (2016), Biochim. Biophys. Acta, 1864, 655-666 .
    View publication on PubMed

Application

EC Number Application Comment Organism
2.4.2.8 drug development the enzyme is a possible target for anti-parasite drug development Trypanosoma cruzi

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.8 gene HGPRTase, recombinant expression of His6-tagged enzyme and of nontagged enzyme in Escherichia coli strain BL21(DE3) Trypanosoma cruzi

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.8 purified recombinant His6-tagged enzyme, mixing of 5 mg/ml protein in 20 mM Tris-HCl, 100 mM NaCl, pH 8.0, with reservoir solution containing 0.1 M MES, pH 6.5, and 12% w/v PEG 20000 at 20°C, X-ray diffraction structure determination and analysis at 2.65 A resolution Trypanosoma cruzi

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.8 additional information truncation of the C-terminal stretch of enzyme TcHPRTH6 enhances its catalytic efficiency Trypanosoma cruzi

General Stability

EC Number General Stability Organism
2.4.2.8 analysis of the stability of the variants in urea-induced unfolding experiments. Protein aggregation is taking place after dialysis or denaturant dilution, an indication of the existence of an irreversible phenomenon, precludes the derivation of thermodynamic state functions from the unfolding data. Nevertheless, the enzyme behaves as cooperative folding units when submitted to chemical denaturation, irreversible aggregation at 60°C within 10 min Trypanosoma cruzi

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.8 Mg2+ required 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 hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate Trypanosoma cruzi
-
 IMP + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.8 Trypanosoma cruzi Q27796
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.8 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis, recombinant nontagged enzyme from Escherichia coli by anion exchange chromatography, ammonium sulfate fractionation, dialysis, and gel filtration, both to over 95% purity Trypanosoma cruzi

Renatured (Commentary)

EC Number Renatured (Comment) Organism
2.4.2.8 although the enzyme protein irreversibly aggregates during unfolding, oligomerization is reversible and can be modulated by low concentrations of urea Trypanosoma cruzi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.8 hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 Trypanosoma cruzi IMP + diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.8 dimer when the C-terminal region, which is predicted as a disordered stretch, is excised by proteolysis, TcHPRT adopts a dimeric state, suggesting that the C-terminal region acts as a main guide for the quaternary arrangement. The C-terminal region exhibits a modulatory role on the enzyme, as attested by the enhanced activity observed for the dimeric form Trypanosoma cruzi
2.4.2.8 tetramer the full-length enzyme form in solution adopts a stable and enzymatically active tetrameric form, exhibiting large intersubunit surfaces, two putative dimeric interfaces Trypanosoma cruzi

Synonyms

EC Number Synonyms Comment Organism
2.4.2.8 HGPRTase
-
 Trypanosoma cruzi
2.4.2.8 hypoxanthine phosphoribosyl transferase
-
 Trypanosoma cruzi
2.4.2.8 TcHPRT
-
 Trypanosoma cruzi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.8 25
-
 assay at Trypanosoma cruzi

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.2.8 60
-
 analysis of the stability of the variants in temperature unfolding experiments. Protein aggregation is taking place after dialysis or denaturant dilution, an indication of the existence of an irreversible phenomenon, precludes the derivation of thermodynamic state functions from the unfolding data. But the enzyme behave as cooperative folding units when submitted to thermal denaturation. In particular, a qualitative comparison (a shift in the observed apparent Tm values) of thermal denaturation curves suggests that the His-tag present in TcHPRTH6 might enhance its thermostability by increasing the height of the activation barrier to the formation of aggregation-prone species. Nevertheless, when the enzyme is incubated for 10 min at 60°C, it irreversibly aggregates with a concomitant loss of enzymatic activity Trypanosoma cruzi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.8 7.4
-
 assay at Trypanosoma cruzi

General Information

EC Number General Information Comment Organism
2.4.2.8 metabolism the metabolism of the protozoan parasite Trypanosoma cruzi depends on the salvage of exogenous purines for nucleotide synthesis. In this context, TcHPRT plays a key role in the survival of trypanosomes in their hosts Trypanosoma cruzi
2.4.2.8 physiological function the hypoxanthine phosphoribosyl transferase (TcHPRT) is a critical enzyme in Trypanosoma cruzi for the survival of the parasite. TcHPRT catalyzes the transfer of a mono phosphorylated ribose from phosphoribosyl diphosphate (PRPP) to the purine ring Trypanosoma cruzi