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Literature summary for 1.14.11.48 extracted from

  • Li, M.; Muller, T.A.; Fraser, B.A.; Hausinger, R.P.
    Characterization of active site variants of xanthine hydroxylase from Aspergillus nidulans (2008), Arch. Biochem. Biophys., 470, 44-53.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutant enzymes Aspergillus nidulans

Protein Variants

Protein Variants Comment Organism
C357A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Mutant enzyme is resistant rto thiol-specific reagents that inactivated wild-type enzyme. Elevated ferroxidase activity in the absence of substrates Aspergillus nidulans
D138A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
D151A inactive mutant enzyme Aspergillus nidulans
E137A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
H149A inactive mutant enzyme Aspergillus nidulans
H340A mutant enzyme exhibits 0.17% of the wild-type enzyme activity Aspergillus nidulans
K122A 2fold increase in the Kd of 2-oxoglutarate compared to wild-type enzyme Aspergillus nidulans
N358A 23fold decrease in kcat/Km compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
Q101A elevated ferroxidase activity in the absence of substrates Aspergillus nidulans
Q356A significant decrease in Ki(app) for 6,8-dihydroxypurine. Elevated ferroxidase activity in the absence of substrates Aspergillus nidulans

Inhibitors

Inhibitors Comment Organism Structure
6,8-dihydroxypurine slow-binding competitive inhibitor Aspergillus nidulans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.031
-
2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
0.032
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
0.041
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
0.042
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
0.045
-
xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
0.047
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
0.048
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
0.053
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
0.057
-
xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
0.058
-
xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
0.059
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
0.069
-
xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
0.071
-
xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
0.088
-
xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
0.122
-
xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
0.4
-
9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
0.55
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
0.554
-
xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
0.58
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
0.68
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
0.78
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
0.88
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.09
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ non-heme mononuclear Fe(II) enzyme Aspergillus nidulans

Organism

Organism UniProt Comment Textmining
Aspergillus nidulans Q4QZZ9
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type enzyme variants are purified from recombinant Escherichia coli cells Aspergillus nidulans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
9-methylxanthine + 2-oxoglutarate + O2
-
Aspergillus nidulans ?
-
?
xanthine + 2-oxoglutarate + O2
-
Aspergillus nidulans urate + succinate + CO2
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Aspergillus nidulans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.66
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.26
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.4
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.9
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
2 8 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
3 6 xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
3.8
-
9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
4.25
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
4.5
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
5.9
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
17
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
19
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
21
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
25
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
31
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
32
-
xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
34
-
xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
35
-
xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
38
-
xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
40
-
xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
42
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
60
-
xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
67
-
2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
72
-
xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Aspergillus nidulans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
70
-
xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
300
-
xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
340
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
360
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
400
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
470
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
490
-
xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
550
-
xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
560
-
xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
610
-
xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
670
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
680
-
xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
760
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
850
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1260
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1310
-
2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1600
-
xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
2050
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
2160
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
2160
-
2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
5410
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
7330
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
8180
-
9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
9500
-
9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans