Cloned (Comment) | Organism |
---|---|
wild-type and mutant enzymes P1G, P1A, Y95F and N97A, expression in Escherichia coli BL21 | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
crystallization of the Y95F mutant enzyme by hanging drop vapor diffusion method, crystal structure of the enzyme with the competitive inhibitor (E)-2-fluoro-p-hydroxycinnamate bound at the active site and that of the protein complexed with the enol form of (p-hydroxyphenyl)pyruvate | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
N97A | the ratio of turnover number to Km-value for enol-phenylpyruvate is 21.3fold higher than that of the wild-type enzyme, the ratio of turnover number to Km-value for enol-(p-hydroxyphenyl)pyruvate is 1.5fold lower than that of the wild-type enzyme, 5fold increase in Ki-value for (E)-2-fluoro-p-hydroxycinnamate compared to the wild-type enzyme | Mus musculus |
P1A | the ratio of turnover number to Km-value for enol-phenylpyruvate is 232fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for enol-(p-hydroxyphenyl)pyruvate is 114fold lower than that of the wild-type enzyme | Mus musculus |
P1G | the ratio of turnover number to Km-value for enol-phenylpyruvate is 232fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for enol-(p-hydroxyphenyl)pyruvate is 143fold lower than that of the wild-type enzyme | Mus musculus |
Y95F | the ratio of turnover number to Km-value for enol-phenylpyruvate is 1.3fold lower than that of the wild-type enzyme, the ratio of turnover number to Km-value for enol-(p-hydroxyphenyl)pyruvate is 1.5fold higher than that of the wild-type enzyme | Mus musculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(E)-2-fluoro-p-hydroxycinnamate | competitive | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.107 | - |
enol-phenylpyruvate | 23°C, mutant enzyme P1G | Mus musculus | |
0.107 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme Y95F | Mus musculus | |
0.11 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme Y97A | Mus musculus | |
0.112 | - |
enol-phenylpyruvate | 23°C, mutant enzyme Y95F | Mus musculus | |
0.143 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme P1G | Mus musculus | |
0.18 | - |
enol-phenylpyruvate | 23°C, mutant enzyme N97A | Mus musculus | |
0.2 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, wild-type enzyme | Mus musculus | |
0.303 | - |
enol-phenylpyruvate | 23°C, mutant enzyme P1A | Mus musculus | |
0.31 | - |
enol-phenylpyruvate | 23°C, wild-type enzyme | Mus musculus | |
0.582 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme P1A | Mus musculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
12335 | - |
x * 12335, electrospray ionization mass spectrometry, mutant enzyme P1G | Mus musculus |
12347 | - |
x * 12347, electrospray ionization mass spectrometry, mutant enzyme N97A | Mus musculus |
12347 | - |
x * 12347, electrospray ionization mass spectrometry, mutant enzyme P1A | Mus musculus |
12358 | - |
x * 12358, electrospray ionization mass spectrometry, mutant enzyme Y95F | Mus musculus |
12371 | - |
x * 12371, electrospray ionization mass spectrometry, wild-type enzyme | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | P34884 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
enol-(p-hydroxyphenyl)pyruvate | - |
Mus musculus | keto-(p-hydroxyphenyl)pyruvate | - |
r | |
enol-phenylpyruvate | - |
Mus musculus | keto-phenylpyruvate | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 12335, electrospray ionization mass spectrometry, mutant enzyme P1G | Mus musculus |
? | x * 12347, electrospray ionization mass spectrometry, mutant enzyme N97A | Mus musculus |
? | x * 12347, electrospray ionization mass spectrometry, mutant enzyme P1A | Mus musculus |
? | x * 12358, electrospray ionization mass spectrometry, mutant enzyme Y95F | Mus musculus |
? | x * 12371, electrospray ionization mass spectrometry, wild-type enzyme | Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
enol-phenylpyruvate | 23°C, mutant enzyme P1G | Mus musculus | |
0.8 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, mutant enzyme P1G | Mus musculus | |
1.7 | - |
enol-phenylpyruvate | 23°C, mutant enzyme P1A | Mus musculus | |
4.1 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme P1A | Mus musculus | |
58 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme N97A | Mus musculus | |
109 | - |
enol-phenylpyruvate | 23°C, mutant enzyme Y95F | Mus musculus | |
126 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, mutant enzyme Y95F | Mus musculus | |
160 | - |
enol-(p-hydroxyphenyl)pyruvate | 23°C, pH 6.5, wild-type enzyme | Mus musculus | |
410 | - |
enol-phenylpyruvate | 23°C, wild-type enzyme | Mus musculus | |
660 | - |
enol-phenylpyruvate | 23°C, mutant enzyme N97A | Mus musculus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
(E)-2-fluoro-p-hydroxycinnamate | 23°C, pH 6.5, mutant enzyme N97A | Mus musculus |