SMALL-SCALE MANUAL MULTIPLE PEPTIDE-SYNTHESIS SYSTEM - APPLICATION TOPHOSPHOPEPTIDE SYNTHESIS

A system for small-scale (ca. 10-50 mumol) manual multiple peptide syn thesis assembled from commercially available solid-phase extraction ap paratus is described. This system was used to prepare (on a 15 mumol s cale) the five monophosphorylated isomers of the peptide ASTTVSKTE, a proteolytic fragment of the C-terminal region of rhodopsin. The peptid es were assembled using serine or threonine active esters without hydr oxyl protection at the positions of phosphorylation. Phosphate groups were introduced using postassembly phosphitylation/oxidation according to a published procedure [Andrews, D.M., Kitchin, J. & Scale, P.W. (1 991) Int. J. Peptide Protein Res. 38, 469-475; Staerkaer, G., Jakobsen , M.H., Olsen, C.E. & Holm, A., Tetrahedron Lett. 32, 5389-5392; Peric h, J.W. (1992) Int. J. Peptide Protein Res. 40 134-140]. The reported system provides a relatively inexpensive approach to multiple peptide synthesis, including synthesis of phosphopeptides, for laboratories wh ose synthesis requirements do not justify investment in an automated m ultiple peptide synthesis instrument. (C) Munksgaard 1993.