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Literature summary for 2.4.2.22 extracted from

  • Ullman, B.; Cyr, N.; Choi, K.; Jardim, A.
    Acidic residues in the purine binding site govern the 6-oxopurine specificity of the Leishmania donovani xanthine phosphoribosyltransferase (2010), Int. J. Biochem. Cell Biol., 42, 253-262.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
Escherichia coli Sphi606 or Sphi609 cells transformed with pSelect-LdXPRT, pSelectldxprtE198D, pSelect-ldxprtE215D, pSelect-ldxprtE215Q and pSelectldxprtE198,215D Leishmania donovani

Protein Variants

Protein Variants Comment Organism
E198D increases the turnover rates of the xanthine phosphoribosyltransferase without altering purine nucleobase specificity, converts the xanthine phosphoribosyltransferase into a broad-specificity enzyme capable of utilizing guanine, hypoxanthine, and xanthine as substrates. Can accommodate GMP in the active site Leishmania donovani
E198D/E215D catalyzes robust guanine phosphoribosylation. Exhibits similar kinetic parameters in the presence of Mg2+ or Mn2+ as the wild-type. Substitution of Mg2+ with Mn2+ does not alter the hypoxanthine phosphoribosylation activity Leishmania donovani
E215D increases the turnover rates of the xanthine phosphoribosyltransferase without altering purine nucleobase specificity, converts the xanthine phosphoribosyltransferase into a broad-specificity enzyme capable of utilizing guanine, hypoxanthine, and xanthine as substrates. Can accommodate GMP in the active site Leishmania donovani
E215Q can accommodate GMP in the active site Leishmania donovani

Inhibitors

Inhibitors Comment Organism Structure
GMP inhibition of the xanthine phosphoribosylation Leishmania donovani

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0021
-
hypoxanthine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0034
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
0.0049
-
guanine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0059
-
diphosphate mutant E198D/E215D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.0073
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
0.0073
-
xanthine wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0078
-
xanthine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0078
-
IMP mutant E198D/E215D, at 25°C, in TM buffer Leishmania donovani
0.008
-
IMP mutant E198D, at 25°C, in TM buffer Leishmania donovani
0.008
-
IMP mutant E215D, at 25°C, in TM buffer Leishmania donovani
0.008
-
IMP mutant E215Q, at 25°C, in TM buffer Leishmania donovani
0.008
-
IMP wild-type, at 25°C, in TM buffer Leishmania donovani
0.0162
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
0.0181
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
0.0199
-
xanthine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.023
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
0.0244
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
0.025
-
guanine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0251
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
0.028
-
diphosphate mutant E215Q, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.0288
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
0.035
-
xanthine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.0382
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
0.039
-
diphosphate wild-type, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.045
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
0.074
-
diphosphate mutant E198D/E215D, in the presence of 8 mM IMP Leishmania donovani
0.0913
-
xanthine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.092
-
9-(5-phospho-beta-D-ribosyl)xanthine wild-type, in TM buffer Leishmania donovani
0.094
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E215Q, in TM buffer Leishmania donovani
0.1
-
guanine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.1
-
guanine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.115
-
diphosphate mutant E215D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.129
-
hypoxanthine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.133
-
diphosphate mutant E198D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.358
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E198D, in TM buffer Leishmania donovani
0.396
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E215D, in TM buffer Leishmania donovani
0.458
-
hypoxanthine wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.524
-
hypoxanthine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
0.533
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E198D/E215D, in TM buffer Leishmania donovani
0.567
-
hypoxanthine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ maximal catalytic activity with 0.06 mM Leishmania donovani
Mn2+ maximal catalytic activity with 0.06 mM. Substitution of Mn2+ for Mg2+ results in ca. 2fold increase in the XPRT turnover rate without altering the xanthine Km value. Replacement of Mg2+ with Mn2+ increases the 5-phospho-alpha-D-ribose 1-diphosphate Km value ca. 2fold. XPRT-catalyzed hypoxanthine phosphoribosylation reveals that substitution of Mg2+ with Mn2+ results in a ca. 4fold increase in the hypoxanthine kcat value and ca. 3fold decrease in the hypoxanthine Km value Leishmania donovani

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
gel filtration Leishmania donovani

Organism

Organism UniProt Comment Textmining
Leishmania donovani
-
-
-

Purification (Commentary)

Purification (Comment) Organism
by gel filtration, more than 95% pure Leishmania donovani

Storage Stability

Storage Stability Organism
-80°C, 10 mM Tris-HCl buffer, pH 7.5 Leishmania donovani

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-phospho-alpha-D-ribose 1-diphosphate + guanine
-
Leishmania donovani GMP + diphosphate
-
?
5-phospho-alpha-D-ribose 1-diphosphate + hypoxanthine
-
Leishmania donovani IMP + diphosphate
-
r
5-phospho-alpha-D-ribose 1-diphosphate + xanthine
-
Leishmania donovani 9-(5-phospho-beta-D-ribosyl)xanthine + diphosphate
-
?
9-(5-phospho-beta-D-ribosyl)xanthine + diphosphate
-
Leishmania donovani 5-phospho-alpha-D-ribose 1-diphosphate + xanthine
-
?
IMP + diphosphate
-
Leishmania donovani 5-phospho-alpha-D-ribose 1-diphosphate + hypoxanthine
-
r

Synonyms

Synonyms Comment Organism
XPRT
-
Leishmania donovani

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00367
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E215D, at 25°C, in TM buffer Leishmania donovani
0.0038
-
diphosphate mutant E215D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.0045
-
diphosphate mutant E198D/E215D, in the presence of 8 mM IMP Leishmania donovani
0.006167
-
IMP mutant E198D/E215D, at 25°C, in TM buffer Leishmania donovani
0.0093
-
diphosphate mutant E198D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.0103
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E198D, at 25°C, in TM buffer Leishmania donovani
0.0118
-
diphosphate mutant E198D/E215D, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.0148
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E198D/E215D, at 25°C, in TM buffer Leishmania donovani
0.0228
-
diphosphate wild-type, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.025
-
diphosphate mutant E215Q, in the presence of 1 mM 9-(5-phospho-beta-D-ribosyl)xanthine Leishmania donovani
0.028
-
9-(5-phospho-beta-D-ribosyl)xanthine wild-type, at 25°C, in TM buffer Leishmania donovani
0.043
-
9-(5-phospho-beta-D-ribosyl)xanthine mutant E215Q, at 25°C, in TM buffer Leishmania donovani
0.1
-
guanine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
1.1
-
guanine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
1.5
-
guanine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
2
-
xanthine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
2.1
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
2.2
-
hypoxanthine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
2.4
-
hypoxanthine mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
2.6
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215Q, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
3.1
-
hypoxanthine wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
3.3
-
guanine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
3.5
-
xanthine wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
3.6
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
3.9
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
3.9
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
6.7
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
7.1
-
hypoxanthine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
8.4
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM hypoxanthine Leishmania donovani
8.7
-
hypoxanthine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
12.3
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
15.1
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
16.4
-
xanthine mutant E198D/E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
17.5
-
xanthine mutant E215D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani
20.7
-
5-phospho-alpha-D-ribose 1-diphosphate mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM xanthine Leishmania donovani
23.3
-
xanthine mutant E198D, at 25°C, 40 mM Tris-HCl, pH 7.5, 1 mM 5-phospho-alpha-D-ribose 1-diphosphate Leishmania donovani

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.039
-
GMP wild-type Leishmania donovani
0.044
-
GMP mutant E215D Leishmania donovani
0.067
-
GMP mutant E198D/E215D Leishmania donovani

General Information

General Information Comment Organism
physiological function Sphi609 cells expressing wild-type XPRT grow in minimal medium supplemented with xanthine but not in medium containing hypoxanthine or guanine. In contrast, Escherichia coli Sphi609 transformed with E198D or E215D exhibit vigorous growth with xanthine or hypoxanthine but notably less growth with guanine. Sphi609 transformants expressing E215Q or E198D/E215D proliferate robustly in xanthine, hypoxanthine, and guanine Leishmania donovani