Literature summary for 3.1.3.48 extracted from

Lountos, G.; Kurussi, S.; Zhao, B.; Dyas, B.; Burke, T.J.; Ulrich, R.; Waugh, D.
High-resolution crystal structures of the D1 and D2 domains of protein tyrosine phosphatase epsilon for structure-based drug design (2018), Acta Crystallogr. Sect. D, 74, 1015-1026 .

Cloned(Commentary)

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 (Commentary)

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

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

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

Inhibitors	Comment	Organism	Structure
additional information	structure-based drug design	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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

Localization	Comment	Organism	GeneOntology No.	Textmining
plasma membrane	-	Homo sapiens	5886	-

Natural Substrates/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Homo sapiens	P23469	-	-

Purification (Commentary)

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

Source Tissue	Comment	Organism	Textmining
breast cancer cell	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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

Synonyms	Comment	Organism
protein tyrosine phosphatase epsilon	-	Homo sapiens
PTPepsilon	-	Homo sapiens
receptor PTPepsilon	-	Homo sapiens
RPTPepsilon	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	25	assay at	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.4	-	assay at	Homo sapiens

General Information

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 [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
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