Pieter Van de Vijver and Dennis Suylen contributed equally to this work.
Nε-(thiaprolyl)-lysine as a handle for site-specific protein conjugation
Article first published online: 30 JUN 2010
Copyright © 2010 Wiley Periodicals, Inc.
Special Issue: Special Issue Dedicated to the 2009 Bruce Merrifield Award Winner Stephen B. H. Kent
Volume 94, Issue 4, pages 465–474, 2010
How to Cite
Van de Vijver, P., Suylen, D., Dirksen, A., Dawson, P. E. and Hackeng, T. M. (2010), Nε-(thiaprolyl)-lysine as a handle for site-specific protein conjugation. Biopolymers, 94: 465–474. doi: 10.1002/bip.21485
- Issue published online: 30 JUN 2010
- Article first published online: 30 JUN 2010
- Manuscript Accepted: 19 APR 2010
- Manuscript Revised: 9 APR 2010
- Manuscript Received: 3 MAR 2010
- synthesized proteins;
- peptide synthesis
In this article, we introduce the use of a thiaproline-modified lysine side-chain [Lys(Thz)], as an unlockable handle that enables late-stage, site-selective modification of chemically synthesized proteins. The Lys(Thz) residue was incorporated into the murine chemokine RANTES to demonstrate its compatibility with Boc/Bzl solid phase peptide synthesis, native chemical ligation, and disulfide bond formation. After oxidative folding of the protein, the thiol was liberated under mild reaction conditions [0.2M hydroxylamine (NH2OH) or O-methylhydroxylamine (MeONH2), pH 4] and was subsequently reacted with thiol-selective tags. This side chain protection strategy enables the use of readily available thiol-reactive probes for the modification of internally disulfide bonded proteins. © 2010 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 94: 465–474, 2010.