Journal of Peptide Research 1997-08-01

The solid-phase synthesis of side-chain-phosphorylated peptide-4-nitroanilides.

A Bernhardt, M Drewello, M Schutkowski

Index: J. Pept. Res. 50(2) , 143-52, (1997)

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Abstract

Peptide-4-nitroanilides can be quickly synthesised using an Fmoc-based approach on 2-chlorotritylchloride resin. Preformed building blocks Fmoc-Xaa-NH-Np (Xaa = Cit, Cys, Gln, His, Lys, Orn, Ser, Thr, Tyr, Trp) can be attached via side chain to the 2-chlorotritylchloride linker of the resin. N-terminal elongation yields the respective peptide-4-nitroanilides after detachment from the solid support. We synthesised a set of tetrapeptide-4-nitroanilides with the general structure Suc-Ala-Phe-Pro-Xaa-NH-Np (Xaa = Asp, Cit, Cys, Glu, Gln, His, Lys, Orn, Ser, Thr, Tyr, Trp). Even peptidyl-arginine-4-nitroanilides are available by a slightly modified procedure. First, the appropriate ornithine-containing peptide was synthesised. After detachment of the peptide from the resin the side-chain primary amino group was transformed to the guanidino function of arginine using 1-guanyl-3,5-dimethylpyrazole. A further application of this method is the convenient synthesis of phosphorylated peptide-4-nitroanilides. Five phosphopeptides with the general structure Ac-Ala-Xaa(PO3H2)-Pro-Yaa-NH-Np (Xaa = Ser, Thr, Tyr; Yaa = Tyr, Lys) and their nonphosphorylated analogues were prepared. Global phosphorylation was carried out on the resin-bound peptides using dibenzyl-N, N-diisopropyl-phosphoramidite/tetrazole followed by oxidation with tert-butyl hydroperoxide.