Proceedings book download - 5Z.com

24 hours. The native configuration of the disulfide core was obtained in 93% purity withinthe crude mixture. Using an orthogonal protecting group strategy, Min-23 could be foldedinto the desired configuration in a two step procedure. The uniformity of both foldedpeptides was verified by HPLC. High resolution mass spectrometry in combination withHPLC enables the identification of the separated peptide species.The Fmoc-assisted solid-phase synthesis and the oxidative peptide folding of thescaffold Min-23 were optimized. In addition, the consecutive oxidation folding strategyallows both a directed peptide folding into the desired disulfide configuration and a controlof the autonomous folding procedure. These achievements of the cystine peptide chemistryare used for engineering random peptide motifs incorporated in the disulfide-constrainedMin-23 scaffold, which are identified by in vitro high-throughput screening techniques.Therefore, this chemical concept is a valuable approach for the miniprotein engineering ofrandomised Min-23 scaffolds.AcknowledgmentsThe authors are grateful to the Bundesministerium für Bildung und Forschung (Grant No. 13N10269)for financial support.References1. Binz, H.K., Amstutz, P., Pluckthun, A. Nat. Biotechnol. 23, 1257 (2005).2. Nygren, P.A., Skerra, A. J. Immunol. Methods 3, 290 (2004).3. Heitz, A., Le-Nguyen, D., Chiche, L. Biochemistry 38, 10615 (1999).4. Souriau, C., Chiche, L., Irving, R., Hudson, P. Biochemistry 44, 7143 (2005).5. Wöhr, T., Wahl, F., Nefzi, A., Rohwedder, B., Sato, T., Sun, X., Mutter, M. J. Am. Chem. Soc. 19,9218 (1996).535