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Hexafluoro-2.propanol as a potent cosolvent for chemical ligation of membrane proteins

science china Chemistry 2011年第1期54卷 110-116页

作者： shen fei,tang {2. & liu lei dep.r.ment of Chemistry,univer.ity of science and technology of china,hefei 2.002.,china key laboratory of bioorganic {12. Chemistry and chemical biology,ministr. of education dep.r.ment of Chemistry,Tsinghua univer.ity.Beijing 100084,china 1. dep.r.ment of Chemistry University of Science and Technology of China Hefei 230026 China2. Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology Ministry of Education dep.r.ment of Chemistry Tsinghua University Beijing 100084 China

The study on membrane proteins is an important challenge mainly because of their very poor solubility in various *** traditional recombinant expression strategy and the native chemical ligation method both have difficulty in generating sufficient amounts of desired proteins with high *** 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 *** both HPLC and UV analyses we concluded that 60% HFIP/8 M urea constituted a good solvent *** 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.

关键词： membrane protein native chemical ligation alternative solvent hexafluoro-2.propanol

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