文章基本信息

标题：A promiscuous split intein with expanded protein engineering applications
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作者：Adam J. Stevens ; Giridhar Sekar ; Neel H. Shah 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2017
卷号：114
期号：32
页码：8538-8543
DOI：10.1073/pnas.1701083114
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The protein trans -splicing (PTS) activity of naturally split inteins has found widespread use in chemical biology and biotechnology. However, currently used naturally split inteins suffer from an “extein dependence,” whereby residues surrounding the splice junction strongly affect splicing efficiency, limiting the general applicability of many PTS-based methods. To address this, we describe a mechanism-guided protein engineering approach that imbues ultrafast DnaE split inteins with minimal extein dependence. The resulting “promiscuous” inteins are shown to be superior reagents for protein cyclization and protein semisynthesis, with the latter illustrated through the modification of native cellular chromatin. The promiscuous inteins reported here thus improve the applicability of existing PTS methods and should enable future efforts to engineer promiscuity into other naturally split inteins.
关键词：chemical biology ; protein engineering ; intein splicing ; protein semisynthesis