Protein-tyrosine phosphatases (PTPs) are vital for regulating tryosine phosphorylation in many processes, including growth and differentiation1,2. The regulation of receptor-like PTP (RPTP) activity remains poorly understood, but based on the crystal structure of RPTPα domain 1 we have proposed that dimerization can negatively regulate activity, through the interaction of an inhibitory ‘wedge’ on one monomer with the catalytic cleft of domain 1 in the other monomer3,4. Here we show that dimerization inhibits the activity of a full-length RPTP in vivo. We generated stable disulphide-bonded full-length RPTPα homodimers by expressing mutants with single cysteines at different positions in the ectodomain juxtamembrane region. Expression of wild-type RPTPα and Phe135Cys and Thr141Cys mutants in RPTPα-null mouse embryo cells increased dephosphorylation and activity of Tyr 529 in the protein tyrosine kinase c-Src; in contrast, expression of a Pro137Cys mutant did not. Mutation of Pro 210/211 to leucine in the inhibitory wedge of the Pro137Cys mutant restored its ability to activate c-Src, indicating that dimerization may inhibit full-length RPTPα activity in a manner stereochemically consistent with RPTPα crystal structures3. Our results suggest that RPTPα activity can in principle be negatively regulated by dimerization in vivo.
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We are grateful to C. Joazeiro for providing the pCR2 retroviral vector and to W. Eckhart and P. Blume-Jensen for suggestions and critical review of the manuscript. This work is supported by grants from the National Cancer Institute (T.H.) and Dutch Cancer Society (J.d.H.). T.H. is a Frank and Else Schilling American Cancer Society Research Professor. G.J. is supported by a fellowship from the American Cancer Society.
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