Cloning and expression of PTP-PEST. A novel, human, nontransmembrane protein tyrosine phosphatase

Yang, Q., Co, D., Sommercorn, J., Tonks, N. K. (March 1993) Cloning and expression of PTP-PEST. A novel, human, nontransmembrane protein tyrosine phosphatase. Journal of Biological Chemistry, 268 (9). pp. 6622-6628. ISSN 0021-9258

URL: http://www.ncbi.nlm.nih.gov/pubmed/8454633

Abstract

The polymerase chain reaction was used to amplify protein tyrosine phosphatase (PTPase)-related cDNA from a template of total RNA isolated from human skeletal muscle. A novel PTPase, which we term PTP-PEST, was detected by this method. The polymerase chain reaction fragment was used to screen two different HeLa cell libraries to obtain full length cDNA clones. The cDNA predicts a protein of 510 amino acids, approximately 60 kDa, that does not contain an obvious signal sequence or transmembrane segment suggesting it is a nonreceptor type enzyme. The PTPase domain is located in the N-terminal portion of the molecule and displays approximately 35% identity to other members of this family of enzymes. The C-terminal segment is rich in Pro, Glu, Asp, Ser, and Thr residues, possessing features of PEST motifs which have previously been identified in proteins with very short intracellular half-lives. The protein was expressed in Escherichia coli as a fusion product with glutathione S-transferase. Intrinsic activity was demonstrated in vitro against a variety of phosphotyrosine-containing substrates including BIRK, the autophosphorylated cytoplasmic kinase domain of the insulin receptor beta subunit. It did not dephosphorylate phosphoseryl-phosphorylase a. PTP-PEST mRNA is broadly distributed in a variety of cell lines. Stimulation of human rhabdomyosarcoma A204 cells, a transformed muscle line, with insulin led to an approximately 4-fold induction of PTP-PEST mRNA within 36 h.

Item Type: Paper
Uncontrolled Keywords: HUMAN-PLACENTA XENOPUS OOCYTES SKELETAL-MUSCLE PURIFICATION INSULIN CELLS SEQUENCE KINASES FAMILY MICROINJECTION
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
Investigative techniques and equipment > cloning
Investigative techniques and equipment > assays > cloning

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > protein tyrosine phosphatase
CSHL Authors:
Communities: CSHL labs > Tonks lab
Depositing User: Matt Covey
Date: March 1993
Date Deposited: 17 Dec 2013 17:01
Last Modified: 17 Dec 2013 17:01
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29072

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