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Synthesis of l ‐ and d ‐Ubiquitin by One‐Pot Ligation and Metal‐Free Desulfurization
Author(s) -
Huang YiChao,
Chen ChenChen,
Gao Shuai,
Wang YeHai,
Xiao Hua,
Wang Feng,
Tian ChangLin,
Li YiMing
Publication year - 2016
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201600101
Subject(s) - ubiquitin , ligation , chemistry , biochemistry , microbiology and biotechnology , biology , gene
Native chemical ligation combined with desulfurization has become a powerful strategy for the chemical synthesis of proteins. Here we describe the use of a new thiol additive, methyl thioglycolate, to accomplish one‐pot native chemical ligation and metal‐free desulfurization for chemical protein synthesis. This one‐pot strategy was used to prepare ubiquitin from two or three peptide segments. Circular dichroism spectroscopy and racemic protein X‐ray crystallography confirmed the correct folding of ubiquitin. Our results demonstrate that proteins synthesized chemically by streamlined 9‐fluorenylmethoxycarbonyl (Fmoc) solid‐phase peptide synthesis coupled with a one‐pot ligation–desulfurization strategy can supply useful molecules with sufficient purity for crystallographic studies.