Chemical synthesis of phosphopeptides using the arylthio group for protection of phosphate: application to identification of cdc2 kinase phosphorylation sites.

Synthesis of two O-phosphorylated dipeptides, N-Boc-O[S,S-bis(p-methoxyphenyl)phosphorodithioyl] -serylproline and N-Boc-O-[S,S-bis(p-methoxyphenyl) phosphorodithioyl]threonylproline, as new O-phosphorylated dipeptide building blocks for the synthesis of O-phosphorylated peptides, is described. Peptides containing phosphoserine or phosphothreonine were prepared by use of these building blocks in the Boc mode of liquid-phase peptide synthesis. Sites phosphorylated by cdc2 kinase were easily identified using these chemically synthesized phosphopeptides.