Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons, expression of the His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in strain DH5alpha | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E187C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
E187C/S188C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
F183C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
G184C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
additional information | development of an engineering method for insertion of a chemical off switch into the catalytically essential loop region, the socalled WPD loop, of a protein tyrosine phosphatase using a combination of point mutations and amino acid insertions, the mutants displays FlAsH-binding cysteine residues, overview. Inhibition of the FlAsH-sensitized TCPTP mutants is rapid and specific, and strong FlAsH sensitivity is observed in mutants that contain as few as two cysteine point mutations in their engineered WPD loops | Homo sapiens |
P181C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
P181C/E187C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
P181C/P186C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
P186C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
P186C/E187C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
V185C | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
FlAsH | the small molecule biarsenical fluorescein derivative, FlAsH, is no inhibitor of the wild-type enzyme but of the engineered WPD insertion mutant enzyme that displays FlAsH-binding cysteine residues. Inhibition of the FlAsH-sensitized TCPTP mutants is rapid and specific, and strong FlAsH sensitivity is observed in mutants that contain as few as two cysteine point mutations in their engineered WPD loops, minimization of FlAsH-binding determinants, overview | Homo sapiens | |
additional information | no inhibition by EDTA at 1 mM | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of WPD loop insertion cysteine mutants, overview | Homo sapiens | |
0.54 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C | Homo sapiens | |
0.85 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/E187C | Homo sapiens | |
1.3 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant G184C | Homo sapiens | |
1.9 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/P186C | Homo sapiens | |
2.3 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C/S188C | Homo sapiens | |
2.5 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, wild-type enzyme | Homo sapiens | |
2.5 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C | Homo sapiens | |
2.7 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C | Homo sapiens | |
2.7 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant V185C | Homo sapiens | |
3.1 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant F183C | Homo sapiens | |
3.1 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C/E187C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P17706 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
T-lymphocyte | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl phosphate + H2O | - |
Homo sapiens | 4-nitrophenol + phosphate | - |
? | |
DADEpYLIPQQG + H2O | a phosphopeptide substrate | Homo sapiens | DADEYLIPQQG + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
protein tyrosine phosphatase | - |
Homo sapiens |
PTP | - |
Homo sapiens |
T-cell PTP | - |
Homo sapiens |
TCPTP | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.83 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant F183C | Homo sapiens | |
1.7 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C/E187C | Homo sapiens | |
3.2 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/P186C | Homo sapiens | |
3.5 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C | Homo sapiens | |
3.7 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C | Homo sapiens | |
3.9 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant V185C | Homo sapiens | |
4.4 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant G184C | Homo sapiens | |
5.1 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, wild-type enzyme | Homo sapiens | |
5.4 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C/S188C | Homo sapiens | |
6.4 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/E187C | Homo sapiens | |
9.8 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics of WPD loop insertion cysteine mutants with FlAsH, overview | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant F183C | Homo sapiens | |
0.53 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C/E187C | Homo sapiens | |
1.3 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C | Homo sapiens | |
1.5 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P186C | Homo sapiens | |
1.5 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant V185C | Homo sapiens | |
1.7 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/P186C | Homo sapiens | |
2.1 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, wild-type enzyme | Homo sapiens | |
2.4 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant E187C/S188C | Homo sapiens | |
3.4 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant G184C | Homo sapiens | |
7.6 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C/E187C | Homo sapiens | |
18 | - |
4-nitrophenyl phosphate | pH 7.0, 22°C, mutant P181C | Homo sapiens |