Expedient Total Synthesis of Small to Medium-Sized Membrane Proteins via Fmoc Chemistry
Expedient Total Synthesis of Small to Medium-Sized Membrane Proteins via Fmoc Chemistry
复制标题
通过 Fmoc 化学便捷地全合成中小型膜蛋白
DOI：
10.1021/ja500222u
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发表时间：
2014-03-05
影响因子：
15
通讯作者：
Liu, Lei
中科院分区：
文献类型：
--
作者：
Zheng, Ji-Shen;Yu, Mu;Liu, Lei
Total chemical synthesis provides a unique approach for the access to uncontaminated, monodisperse, and more importantly, post-translationally modified membrane proteins. In the present study we report a practical procedure for expedient and cost-effective synthesis of small to medium-sized membrane proteins in multimilligram scale through the use of automated Fmoc chemistry. The key finding of our study is that after the attachment of a removable arginine-tagged backbone modification group, the membrane protein segments behave almost the same as ordinary water-soluble peptides in terms of Fmoc solid-phase synthesis, ligation, purification, and mass spectrometry characterization. The efficiency and practicality of the new method is demonstrated by the successful preparation of Ser64-phosphorylated M2 proton channel from influenza A virus and the membrane-embedded domain of an inward rectifier K+ channel protein Kir5.1. Functional characterizations of these chemically synthesized membrane proteins indicate that they provide useful and otherwise-difficult-to-access materials for biochemistry and biophysics studies.
