Cloned (Comment) | Organism |
---|---|
recombinant expression of the TEV protease-cleavable, N-terminally His6-tagged maltose-binding protein-PTPepsilon D1(Ser107-Gly398) fusion protein and of wild-type and mutant TEV protease-cleavable, N-terminally His6-tagged-maltose-binding protein D2 domain of PTPepsilon D2(Gly425-Lys700) in Escherichia coli strain Rosetta 2 (DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type and mutant isolated membrane-proximal D1 and distal D2 domains of protein tyrosine phosphatase epsilon, and PTPepsilon (A455N/V457Y/E597D) D2 domain mutant, hanging drop vapour diffusion method, for PTPepsilon D1 domain mixing of 0.002 ml of 13.9 mg/ml protein solution with 0.002 ml of reservoir solution containing 0.1 M Tris-HCl, pH 8.5, 0.2 M sodium chloride, and 22% w/v PEG 3350, for wild-type PTPepsilon D2 domain mixing of 0.001 ml of 5.0 mg/ml protein solution with 0.003 ml of reservoir solution containing 0.1 M bicine/trizma base, pH 8.5, 0.03 mM diethylene glycol, 0.03 M triethylene glycol, 0.03 M tetraethylene glycol, 0.03 M pentaethylene glycol, 10% w/v PEG 4000, and 20% v/v glycerol, and for mutant A455N/V457Y/E597D PTPepsilon D2 domain mixing of 0.002 ml of 9.0 mg/ml protein solution with 0.002 ml of reservoir solution containing 0.1 M imidazole, pH 8.0, 0.2 M calcium acetate hydrate, 20% w/v PEG 1000, in all cases equilibration against 0.5 ml reservoir solution, X-ray diffraction structure determination and analysis at 1.67-2.27 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A455N/V457Y/E597D | site-directed mutagenesis, the triple mutant of the PTPepsilon D2 domain is constructed to reconstitute the residues of the PTPepsilon D1 catalytic domain that are important for phosphatase activity, resulting in only a slight increase in the phosphatase activity compared with the wild-type D2 protein. The mutant is used for structure-based drug design | Homo sapiens |
V457Y/E597D | site-directed mutagenesis | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | structure-based drug design | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.119 | - |
PQDKEY-pY-KVKEPG | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a JAK2 peptide | Homo sapiens | |
0.124 | - |
RKGSGD-pY-MPMSPK | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a IRS1 peptide | Homo sapiens | |
0.127 | - |
PGSAAP-pY-LKTKFI | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a STAT3 peptide | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
plasma membrane | - |
Homo sapiens | 5886 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
[a protein]-tyrosine phosphate + H2O | Homo sapiens | - |
[a protein]-tyrosine + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P23469 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant TEV protease-cleavable, N-terminally His6-tagged maltose-binding protein-PTPepsilon D1 and D2 fusion proteins from Escherichia coli strain Rosetta 2 (DE3) by nickel affinity chromatography, ultrafiltration, and cleavage of the tag/fusion protein, followed by gel filtration and ultrafiltration | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
breast cancer cell | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | activity detection by malachite green phosphatase method for phosphate determination | Homo sapiens | ? | - |
? | |
PGSAAP-pY-LKTKFI + H2O | STAT3 peptide | Homo sapiens | PGSAAPYLKTKFI + phosphate | - |
? | |
PQDKEY-pY-KVKEPG + H2O | JAK2 peptide | Homo sapiens | PQDKEYYKVKEPG + phosphate | - |
? | |
RKGSGD-pY-MPMSPK + H2O | IRS1 peptide | Homo sapiens | RKGSGDYMPMSPK + phosphate | - |
? | |
[a protein]-tyrosine phosphate + H2O | - |
Homo sapiens | [a protein]-tyrosine + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the receptor PTPepsilon is located in the plasma membrane and possesses a small extracellular domain, a membrane-spanning domain and two tandem cytoplasmic domains, D1 and D2 | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
protein tyrosine phosphatase epsilon | - |
Homo sapiens |
PTPepsilon | - |
Homo sapiens |
receptor PTPepsilon | - |
Homo sapiens |
RPTPepsilon | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | 25 | assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.1 | - |
PQDKEY-pY-KVKEPG | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a JAK2 peptide | Homo sapiens | |
3.2 | - |
RKGSGD-pY-MPMSPK | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a IRS1 peptide | Homo sapiens | |
3.6 | - |
PGSAAP-pY-LKTKFI | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a STAT3 peptide | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.4 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
additional information | PTPepsilon D1 and D2 active site structures, overview | Homo sapiens |
physiological function | the reversible phosphorylation of tyrosine residues in proteins, which depends on the balanced coordination between a multitude of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs), is an essential step in many cellular signaling pathways. Classical type I cysteine-based PTPs include both receptor and nonreceptor forms that catalyze the removal of the phosphate moiety from a targeted pTyr residue. The enzyme plays a positive role in the survival of human breast cancer cells | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
25.81 | - |
RKGSGD-pY-MPMSPK | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a IRS1 peptide | Homo sapiens | |
26.05 | - |
PQDKEY-pY-KVKEPG | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a JAK2 peptide | Homo sapiens | |
28.35 | - |
PGSAAP-pY-LKTKFI | pH 6.4, 25°C, recombinant PTPepsilon D1 domain, substrate is a STAT3 peptide | Homo sapiens |