Efficient Fmoc/solid‐phase peptide synthesis of O‐phosphotyrosyl‐containing peptides and their use as phosphatase substrates

Abstract

A general synthetic method for the efficient preparation of Tyr(P) ‐containing peptides is described by the use of Fmoc‐Tyr(PO31Bu2) ‐OH in Fmoc/solid‐phase synthesis followed by simultaneous cleavage of the peptide from the resin and peptide deprotection by acidolytic treatment. The applicability of this approach is demonstrated by the synthesis of H‐Ser‐Ser‐Ser‐Tyr(P) ‐Tyr(P) ‐OH.TFA and the synthesis of the phosphorylated forms of the two physiological peptides, angiotensin II and neurotensin 8–13. In addition, the three phosphorylated peptides were used as substrates in the study of the local specificity determinants of T‐cell protein tyrosine phosphatase. In a competition assay using 32..