Hexafluoro-2-propanol as a potent cosolvent for chemical ligation of membrane proteins
Hexafluoro-2-propanol as a potent cosolvent for chemical ligation of membrane proteins
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六氟-2-丙醇作为膜蛋白化学连接的有效共溶剂
DOI：
10.1007/s11426-010-4188-4
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发表时间：
2011-01
期刊：
影响因子：
--
通讯作者：
Department of Chemistry,Tsinghua University.Beijin
中科院分区：
文献类型：
--
作者：
SHEN Fei,TANG Shan & LIU Lei Department of Chemist;Department of Chemistry,Tsinghua University.Beijin
膜蛋白的研究是一个重要的挑战，主要是因为它们在各种溶剂中的溶解性很差。传统的重组表达策略和天然的化学连接方法都难以高效地产生足够数量的目的蛋白。以往的研究表明，多元氟化醇对难溶的多肽序列表现出很好的溶解能力，尤其是六氟丙醇(HFIP)。在本研究中，我们系统地研究了含有不同比例的HFIP的溶剂对跨膜肽的溶解能力。通过高效液相色谱和紫外光谱分析，我们认为60%HFIP/8M尿素是一个很好的溶剂体系。在这个溶剂体系中，我们还优化了进行天然化学连接(NCL)的条件。在优化的条件下，我们成功地实现了NCL对二肽的形成和模型蛋白(Trifolitoxin)的合成。这些结果表明，HFIP是一种潜在的助溶剂，可用于连接难溶性多肽以产生膜蛋白。
The study on membrane proteins is an important challenge mainly because of their very poor solubility in various solvents. The traditional recombinant expression strategy and the native chemical ligation method both have difficulty in generating sufficient amounts of desired proteins with high efficiency. Previous studies have shown that multiply fluorinated alcohols exhibit good ability to dissolve difficult peptide sequences, especially hexafluoro-2-propanol (HFIP). In the present study we systematically studied the capability of solvents containing different percentage of HFIP in dissolving transmembrane peptides. Through both HPLC and UV analyses we concluded that 60% HFIP/8 M urea constituted a good solvent system. In this solvent system we also optimized conditions to perform native chemical ligation (NCL). Under the optimized conditions we successfully achieved NCL’s for both dipeptide formation and the synthesis of a model protein (Trifolitoxin). These results suggested that HFIP was a potential cosolvent that could be used in the ligation of poorly soluble peptides for the generation of membrane proteins.
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