期刊
CHEMISTRY-A EUROPEAN JOURNAL
卷 22, 期 22, 页码 7623-7628出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/chem.201600101
关键词
desulfurization; methyl thioglycolate; native chemical ligation; proteins; X-ray diffraction
资金
- NSFC [21372058, 21572043, 21302034]
- State Key Laboratory of Medicinal Chemical Biology
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.
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