Synthesis of Human Interleukin-8 by Native Chemical Ligation

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Synthesis of Human Interleukin-8 by Native Chemical Ligation

P. E. DAWSON, T. W. MUIR, I. CLARK-LEWIS, S. B. KENT* (THE SCRIPPS RESEARCH Category INSTITUTE, LA JOLLA, USA) Chemistry in Synthesis of Proteins by Native Chemical Ligation Medicine and Science 1994, 266, 776–779. Biology Synthesis of Human Interleukin-8 by Native Chemical Key words Ligation peptide synthesis native chemical ligation Principle of native chemical ligation (NCL): protein synthesis O O human interleukin 8 Unprotected peptide H3N peptide-1 SR peptide-2 segments C-terminal thioester N-terminal cysteine S chemoselective thioesterification O O H2N Intermediate thioester peptide-1 peptide-2 linkage S spontaneous amide bond formation O O Final ligated product with peptide-1 N peptide-2 native amide bond H HS Synthesis of human interleukin 8 (IL-8) via NCL: O O O H N 3 – H3N IL-8 (1-33) SBn + IL-8 (35-72) O HS SH HS SH ⋅ 6 M guanidine HCl, BnSH ca. 61% yield peptide fragments pH 7.6 phosphate buffer obtained by SPPS O O O – H3N IL-8 (1-33) N IL-8 (35-72) O H HS SH HS SH air, 1 M guanidine⋅HCl folding and disulfide formation pH 8.5 tris buffer O O O – H3N IL-8 (1-33) N IL-8 (35-72) O H S S This document was downloaded for personal use only. Unauthorized distribution is strictly prohibited. S S IL-8 Significance: The realization that unprotected Comment: NCL proceeds by the formation of an peptide thioesters react chemoselectively with un- intermediate thioester linkage with cysteine that protected N-terminal cysteine residues to form am- then spontaneously rearranges to the native amide ide linkages is an important advancement in pro- bond. This strategy was applied to the total chemi- tein synthesis. Native Chemical Ligation (NCL) cal synthesis of human interleukin 8 (IL-8), a 72- allows synthetic access to proteins of moderate amino acid protein containing four cysteine resi- size. dues. SYNFACTS Contributors: Dirk Trauner, Julius R. Reyes Synfacts 02042019, 15(04), 0439 Published online: 19.03.20191861-19581861-194X DOI: 10.1055/s-0037-1612404; Reg-No.: T01919SF ©Georg Thieme Verlag Stutgart · New York 2019 © Georg Thieme Verlag Stuttgart · New York 439.

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