The stapling systems described in this protocol, namely bridging one or two turns of an -helix, are highly adaptable to most peptide sequences, resulting in favorable RCM kinetics, helix stabilization and promotion of cellular uptake. The nomenclature R i, i +3 S (8) refers to an eight-carbon metathesized cross-link with R -configuration at i and S -configuration at i +3 position; S i, i +4 S (8), 8-carbon tether with S -configuration at both i and i +4 positions; R i, i +7 S (11), 11-carbon tether with R -configuration at i and S -configuration at i +7 position. ( a ) Olefin-bearing non-natural amino Full Report acids used for the synthesis of hydrocarbon stapled peptides; ( b ) Stapled peptides (SAHBs) were derived from BID BH3 domain. Rcm ring closing metathesis. All authors discussed the results and wrote the manuscript.
18 June 2017 23 June Cover letter admissions counselor position 2017 08 March 2017 09 March 2017 Vienna, Austria 14 July 2017 15 July 2017 Vienna, AustriaNature Protocols 15 Mar 2012Nature Protocols 28 Oct 2010Nature Protocols 15 Sep 2016 Directed the research and, with Y. -W. K., conceptualized the experiments. Performed the experiments. Residues of the BH3 helix that do not participate in direct interactions with the target protein are highlighted in red (N L:
Y. -W. K. Norleucine). After ring-closing olefin metathesis (RCM), the resin-bound peptide can be further modified on its N terminus or side chains, e.
g., by acetylation, fluoresceination or biotinylation. Two product peaks represent olefin isomers. G. L. V. Article previewThis protocol provides a detailed procedure for the preparation of stapled -helical peptides, which have proven their potential as useful molecular probes and as next-generation therapeutics. Two crucial features of this protocol are (i) the construction of peptide substrates containing hindered -methyl, -alkenyl amino acids and (ii) the ring-closing olefin metathesis (RCM) of the resulting resin-bound peptide substrates.