show The role of tyrosine 343 in substrate binding and catalysis by human sulfite oxidase
Wilson, H.L.; Rajagopalan, K.V.; J. Biol. Chem. 279, 15105-15113 (2004)
Data extracted from this reference:
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.8.3.1
Y343F
in the mutant enzyme using cytochrome c as electron acceptor, turnover number is somewhat impaired, 34% of the wild-type activity at pH 8.5. The KM-value for the mutant enzyme shows a 5fold increase over wild-type. Reduction of the molybdenum center of the Y343 F variant by sulfite is more significantly impaired at high pH than at low pH
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.8.3.1
0.00129
-
sulfite
25°C, pH 6.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00162
-
sulfite
25°C, pH 6.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00272
-
sulfite
25°C, pH 7.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00311
-
sulfite
25°C, pH 6.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00339
-
sulfite
25°C, pH 7.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00352
-
sulfite
25°C, pH 7.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00362
-
sulfite
25°C, pH 7.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00367
-
sulfite
25°C, pH 7.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00414
-
sulfite
25°C, pH 6.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00423
-
sulfite
25°C, pH 7.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00435
-
sulfite
25°C, pH 8.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00459
-
sulfite
25°C, pH 7.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00503
-
sulfite
25°C, pH 8.25, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00612
-
sulfite
25°C, pH 8.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00635
-
sulfite
25°C, pH 7.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00728
-
sulfite
25°C, pH 8.25, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00825
-
sulfite
25°C, pH 8.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00851
-
sulfite
25°C, pH 10.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00859
-
sulfite
25°C, pH 8.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00908
-
sulfite
25°C, pH 8.25, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00924
-
sulfite
25°C, pH 8.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00947
-
sulfite
25°C, pH 9.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00959
-
sulfite
25°C, pH 8.75, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00963
-
sulfite
25°C, pH 9.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00992
-
sulfite
25°C, pH 9.75, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.011
-
sulfite
25°C, pH 8.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0158
-
sulfite
25°C, pH 8.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0221
-
sulfite
25°C, pH 9.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.026
-
sulfite
25°C, pH 9.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0319
-
sulfite
25°C, pH 8.25, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0536
-
sulfite
25°C, pH 9.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0557
-
sulfite
25°C, pH 8.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0671
-
sulfite
25°C, pH 9.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.147
-
sulfite
25°C, pH 9.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.59
1
sulfite
25°C, pH 9.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
1.54
-
sulfite
25°C, pH 10.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.8.3.1
Homo sapiens
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.8.3.1
sulfite + O2
-
659319
Homo sapiens
sulfate + H2O2
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.3.1
2.06
-
sulfite
25°C, pH 7.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
3.11
-
sulfite
25°C, pH 6.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
3.26
-
sulfite
25°C, pH 7.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
4.14
-
sulfite
25°C, pH 6.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
4.59
-
sulfite
25°C, pH 7.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
6.35
-
sulfite
25°C, pH 7.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
7.17
-
sulfite
25°C, pH 8.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.21
-
sulfite
25°C, pH 10.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.51
-
sulfite
25°C, pH 10.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.59
-
sulfite
25°C, pH 8.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.72
-
sulfite
25°C, pH 8.25, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.08
-
sulfite
25°C, pH 8.25, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.2
-
sulfite
25°C, pH 8.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.24
-
sulfite
25°C, pH 8.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.47
-
sulfite
25°C, pH 9.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.63
-
sulfite
25°C, pH 9.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.92
-
sulfite
25°C, pH 9.75, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.99
-
sulfite
25°C, pH 9.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
10.5
-
sulfite
25°C, pH 9.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
12.1
-
sulfite
25°C, pH 7.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
13.2
-
sulfite
25°C, pH 6.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
17.2
-
sulfite
25°C, pH 7.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
17.7
-
sulfite
25°C, pH 6.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
23.6
-
sulfite
25°C, pH 8.75, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.2
-
sulfite
25°C, pH 7.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.6
-
sulfite
25°C, pH 9.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.7
-
sulfite
25°C, pH 7.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.8
-
sulfite
25°C, pH 8.25, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25
-
sulfite
25°C, pH 8.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25.7
-
sulfite
25°C, pH 9.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25.9
-
sulfite
25°C, pH 8.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
26.3
-
sulfite
25°C, pH 9.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
26.9
-
sulfite
25°C, pH 8.5, 100 mM buffer, wild-type enzyme; 25°C, pH 8.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
27
-
sulfite
25°C, pH 8.25, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
28.1
-
sulfite
25°C, pH 9.0, 100 mM buffer, wild-type enzyme
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.8.3.1
Y343F
in the mutant enzyme using cytochrome c as electron acceptor, turnover number is somewhat impaired, 34% of the wild-type activity at pH 8.5. The KM-value for the mutant enzyme shows a 5fold increase over wild-type. Reduction of the molybdenum center of the Y343 F variant by sulfite is more significantly impaired at high pH than at low pH
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.8.3.1
0.00129
-
sulfite
25°C, pH 6.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00162
-
sulfite
25°C, pH 6.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00272
-
sulfite
25°C, pH 7.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00311
-
sulfite
25°C, pH 6.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00339
-
sulfite
25°C, pH 7.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00352
-
sulfite
25°C, pH 7.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00362
-
sulfite
25°C, pH 7.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00367
-
sulfite
25°C, pH 7.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00414
-
sulfite
25°C, pH 6.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00423
-
sulfite
25°C, pH 7.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00435
-
sulfite
25°C, pH 8.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00459
-
sulfite
25°C, pH 7.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00503
-
sulfite
25°C, pH 8.25, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00612
-
sulfite
25°C, pH 8.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00635
-
sulfite
25°C, pH 7.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00728
-
sulfite
25°C, pH 8.25, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00825
-
sulfite
25°C, pH 8.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00851
-
sulfite
25°C, pH 10.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00859
-
sulfite
25°C, pH 8.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00908
-
sulfite
25°C, pH 8.25, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00924
-
sulfite
25°C, pH 8.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00947
-
sulfite
25°C, pH 9.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00959
-
sulfite
25°C, pH 8.75, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.00963
-
sulfite
25°C, pH 9.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.00992
-
sulfite
25°C, pH 9.75, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.011
-
sulfite
25°C, pH 8.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0158
-
sulfite
25°C, pH 8.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0221
-
sulfite
25°C, pH 9.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.026
-
sulfite
25°C, pH 9.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0319
-
sulfite
25°C, pH 8.25, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0536
-
sulfite
25°C, pH 9.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.0557
-
sulfite
25°C, pH 8.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.0671
-
sulfite
25°C, pH 9.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
0.147
-
sulfite
25°C, pH 9.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
0.59
1
sulfite
25°C, pH 9.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
1.54
-
sulfite
25°C, pH 10.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.8.3.1
sulfite + O2
-
659319
Homo sapiens
sulfate + H2O2
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.3.1
2.06
-
sulfite
25°C, pH 7.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
3.11
-
sulfite
25°C, pH 6.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
3.26
-
sulfite
25°C, pH 7.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
4.14
-
sulfite
25°C, pH 6.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
4.59
-
sulfite
25°C, pH 7.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
6.35
-
sulfite
25°C, pH 7.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
7.17
-
sulfite
25°C, pH 8.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.21
-
sulfite
25°C, pH 10.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.51
-
sulfite
25°C, pH 10.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.59
-
sulfite
25°C, pH 8.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
8.72
-
sulfite
25°C, pH 8.25, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.08
-
sulfite
25°C, pH 8.25, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.2
-
sulfite
25°C, pH 8.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.24
-
sulfite
25°C, pH 8.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.47
-
sulfite
25°C, pH 9.0, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.63
-
sulfite
25°C, pH 9.5, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.92
-
sulfite
25°C, pH 9.75, 20 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
9.99
-
sulfite
25°C, pH 9.0, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
10.5
-
sulfite
25°C, pH 9.5, 100 mM buffer, mutant enzyme Y343F
Homo sapiens
1.8.3.1
12.1
-
sulfite
25°C, pH 7.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
13.2
-
sulfite
25°C, pH 6.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
17.2
-
sulfite
25°C, pH 7.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
17.7
-
sulfite
25°C, pH 6.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
23.6
-
sulfite
25°C, pH 8.75, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.2
-
sulfite
25°C, pH 7.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.6
-
sulfite
25°C, pH 9.5, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.7
-
sulfite
25°C, pH 7.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
24.8
-
sulfite
25°C, pH 8.25, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25
-
sulfite
25°C, pH 8.0, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25.7
-
sulfite
25°C, pH 9.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
25.9
-
sulfite
25°C, pH 8.0, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
26.3
-
sulfite
25°C, pH 9.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
26.9
-
sulfite
25°C, pH 8.5, 100 mM buffer, wild-type enzyme; 25°C, pH 8.5, 20 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
27
-
sulfite
25°C, pH 8.25, 100 mM buffer, wild-type enzyme
Homo sapiens
1.8.3.1
28.1
-
sulfite
25°C, pH 9.0, 100 mM buffer, wild-type enzyme
Homo sapiens