Evidence for Reversible Tyrosine Protein Phosphorylation in the Okadaic Acid-Producing Marine Dinoflagellate Prorocentrum lima

ABSTRACT. The protist Prorocentrum lima , a primary producer of the tumour promoter okadaic acid, is a member of the dinoflagellate class of marine microorganisms. Herein, we have identified and characterized a protein tyrosine kinase (designated PLIK 1A) in P. lima that autophosphorylates almost exclusively on tyrosine residues. PLIK 1A was shown to have an approximate molecular mass of 38 kDa by SDS-PAGE and a native molecular mass within the range of 47–55 kDa by Superdex-75 gel filtration. Phosphoamino acid analysis of autophosphorylated PLIK 1A revealed the presence of phosphotyrosine and autophosphorylated PLJK 1A reacted with monoclonal anti-phosphotyrosine antibodies in a Western immunoblot. In addition, two protein tyrosine phosphatases were identified in P. lima that had apparent molecular masses within the ranges of 150–168 kDa and 73–82 kDa as determined by Superdex-200 gel filtration. These P. lima phosphatases, termed PLPTP-I and PLPTP-II, efficiently dephosphorylated tyrosine phosphorylated myelin basic protein. owever, only PLPTP-I was capable of dephosphorylating the tyrosine phosphorylated substrate angiotensin. Both PLPTP-I and PLPTP-II were able to dephosphorylate tyrosine autophosphorylated PLIK 1A. These data provide the first evidence for reversible tyrosine protein phosphorylation in P. lima by protein tyrosine kinases and phosphatases