Regulation of protein-tyrosine phosphatases alpha and epsilon by calpain-mediated proteolytic cleavage.
The precise subcellular localization of non-receptor tyrosine phosphatases is a major factor in regulating their physiological functions. We have previously shown that cellular processing of protein-tyrosine phosphatase epsilon (PTP epsilon) generates a physiologically distinct, cytoplasmic form of this protein, p65 PTP epsilon. Here we describe a novel protein form of the related receptor-type tyrosine phosphatase alpha (RPTP alpha), p66 PTP alpha, which is detected in nearly all cell types where RPTP alpha is expressed. Both p66 PTP alpha and p65 PTP epsilon are produced by calpain-mediated proteolytic cleavage in vivo. Cleavage is inhibited in living cells by a variety of calpain inhibitors, can be induced in primary cortical neurons treated with calcium chloride, and is observed in lysates of brain or of cultured cells following addition of purified calpain. Cleavage occurs within the intracellular juxtamembrane domain of RPTP alpha, releasing the phosphatase catalytic domains from their membranal anchors and translocating them to the cytoplasm. Translocation reduces the ability of PTPalpha to act on membrane-associated substrates, as it loses its ability to dephosphorylate Src at its C-terminal regulatory site, and its ability to dephosphorylate the Kv2.1 voltage-gated potassium channel is severely impaired. In all, the data indicate that control of phosphatase function via post-translational processing occurs also among receptor-type phosphatases, and demonstrate the molecular complexity of regulating these parameters within the PTP alpha/PTP epsilon phosphatase subfamily.
Gil-Henn
H
H
Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot 76100, Israel.
Volohonsky
G
G
Elson
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002154:Calpain / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D017931:DNA Primers; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism*; D021381:Protein Transport; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D007527:Isoenzymes; D017027:Protein Tyrosine Phosphatases; D002154:Calpain