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ERK1/2 phosphorylate RSPH3. A, phosphoamino acid analysis of His6-RSP3 phosphorylated in vitro with pERK1. An autoradiogram is shown. B, 3xFLAG-RSP3H wild-type or T286A was immunoprecipitated from HEK293 cells and used as substrate for an in vitro kinase reaction with purified, recombinant pERK1. Mutating T286A decreased 32P incorporation by ∼70% (n = 6). pS, phosphoserine; pT, phosphothreonine; pY, phosphotyrosine.
