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Literature summary for 2.4.2.8 extracted from

  • Keough, D.T.; Ng, A.L.; Winzor, D.J.; Emmerson, B.T.; de Jersey, J.
    Purification and characterization of Plasmodium falciparum hypoxanthine-guanine-xanthine phosphoribosyltransferase and comparison with the human enzyme (1999), Mol. Biochem. Parasitol., 98, 29-41.
    View publication on PubMed

Application

Application Comment Organism
medicine potential target for antiparasitic chemotherapy Homo sapiens
medicine potential target for antiparasitic chemotherapy Plasmodium falciparum

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in an enzyme-deficient Escherichia coli strain Plasmodium falciparum
overexpression in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
C105A prepared via splicing by overlap extension, reduced oxidation ofthe enzyme during storage Homo sapiens
C205A prepared via splicing by overlap extension, reduced oxidation ofthe enzyme during storage Homo sapiens
C22A prepared via splicing by overlap extension, reduced oxidation ofthe enzyme during storage Homo sapiens

General Stability

General Stability Organism
DTT and hypoxanthine stabilize Plasmodium falciparum

Inhibitors

Inhibitors Comment Organism Structure
Hg2+ complete inhibition at 3 mM after 3 min at 0°C Homo sapiens
Hg2+ complete inhibition at 3 mM after 3 min at 0°C Plasmodium falciparum
KCl no inhibition Homo sapiens
KCl inactivation Plasmodium falciparum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0009
-
hypoxanthine recombinant enzyme, with other purine Plasmodium falciparum
0.0014
-
guanine recombinant enzyme, with other purine Plasmodium falciparum
0.0019
-
guanine recombinant enzyme, with other purine Homo sapiens
0.0031
-
hypoxanthine recombinant enzyme, with other purine Homo sapiens
0.0117
-
allopurinol recombinant enzyme Plasmodium falciparum
0.06
-
hypoxanthine recombinant enzyme, with 5-phosphoribosyl 1-diphosphate Plasmodium falciparum
0.126
-
hypoxanthine recombinant enzyme, with 5-phosphoribosyl 1-diphosphate, pH 8.5 Homo sapiens
0.134
-
guanine recombinant enzyme, with 5-phosphoribosyl 1-diphosphate, pH 8.5 Homo sapiens
0.135
-
allopurinol recombinant enzyme Homo sapiens
0.17
-
xanthine recombinant enzyme, with 5-phosphoribosyl 1-diphosphate, pH 8.5 Plasmodium falciparum
0.362
-
guanine recombinant enzyme, with 5-phosphoribosyl 1-diphosphate Plasmodium falciparum
0.42
-
xanthine recombinant enzyme, pH 8.5 Plasmodium falciparum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Homo sapiens
Mg2+
-
Plasmodium falciparum

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Plasmodium falciparum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant from Escherichia coli Homo sapiens
recombinant from Escherichia coli , large scale Plasmodium falciparum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.27
-
purified recombinant enzyme, substrate allopurinol, pH 8.0 Homo sapiens
1.4
-
purified recombinant enzyme, substrate hypoxanthine, pH 8.0 Plasmodium falciparum
3
-
purified recombinant enzyme, substrates allopurinol and xanthine, pH 8.0 Plasmodium falciparum
5.4
-
purified recombinant enzyme, substrate guanine, pH 8.0 Plasmodium falciparum
27
-
purified recombinant enzyme, substrate hypoxanthine, pH 8.0 Homo sapiens
46
-
purified recombinant enzyme, substrate guanine, pH 8.0 Homo sapiens

Storage Stability

Storage Stability Organism
0°C, 10 mM phosphate, pH 7.1, 1 mM DTT, 10 mM Mg2+, 2 h, no loss of activity Homo sapiens
4°C, freshly purified enzyme, 10 mM phosphate, pH 6.8, 10 mM DTT, 1 mM 5-phosphoribosyl 1-diphosphate, rapid loss of 90% activity within 48 h Plasmodium falciparum
5°C, purified enzyme, 10 mM phosphate, pH 6.8, 1 mM DTT, 0.2 mM 5-phosphoribosyl 1-diphosphate, 0.06 hypoxanthine, loss of 7% activity after 4-8 weeks Plasmodium falciparum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
allopurinol + 5-phospho-alpha-D-ribose 1-diphosphate
-
Plasmodium falciparum allopurinol ribonucleoside 5'-monophosphate + diphosphate
-
?
allopurinol + 5-phospho-alpha-D-ribose 1-diphosphate low activity Homo sapiens allopurinol ribonucleoside 5'-monophosphate + diphosphate
-
?
guanine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Homo sapiens GMP + diphosphate
-
?
guanine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Plasmodium falciparum GMP + diphosphate
-
?
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Homo sapiens IMP + diphosphate
-
?
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Plasmodium falciparum IMP + diphosphate
-
?
xanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Homo sapiens XMP + diphosphate
-
?
xanthine + 5-phospho-alpha-D-ribose 1-diphosphate
-
Plasmodium falciparum XMP + diphosphate
-
?

Subunits

Subunits Comment Organism
? x * 24352-24353, recombinant mutant enzymes, mass spectroscopy and DNA sequence determination Homo sapiens
dimer
-
Homo sapiens
dimer 2 * 26229-26232, in presence of KCl, SDS-PAGE, mass spectroscopy and DNA sequence determination Plasmodium falciparum
tetramer
-
Homo sapiens
tetramer 4 * 26229-26232, in absence of KCl, SDS-PAGE, mass spectroscopy and DNA sequence determination Plasmodium falciparum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Plasmodium falciparum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Plasmodium falciparum