Arachea Buenafe T, Sun Zhen, Potente Nina, Malik Radhika, Isailovic Dragan, Viola Ronald E
Department of Chemistry, University of Toledo, 2801 W. Bancroft St., Toledo, OH 43606, United States.
Protein Expr Purif. 2012 Nov;86(1):12-20. doi: 10.1016/j.pep.2012.08.016. Epub 2012 Aug 31.
Generating stable conditions for membrane proteins after extraction from their lipid bilayer environment is essential for subsequent characterization. Detergents are the most widely used means to obtain this stable environment; however, different types of membrane proteins have been found to require detergents with varying properties for optimal extraction efficiency and stability after extraction. The extraction profiles of several detergent types have been examined for membranes isolated from bacteria and yeast, and for a set of recombinant target proteins. The extraction efficiencies of these detergents increase at higher concentrations, and were shown to correlate with their respective CMC values. Two alkyl sugar detergents, octyl-β-d-glucoside (OG) and 5-cyclohexyl-1-pentyl-β-d-maltoside (Cymal-5), and a zwitterionic surfactant, N-decylphosphocholine (Fos-choline-10), were generally effective in the extraction of a broad range of membrane proteins. However, certain detergents were more effective than others in the extraction of specific classes of integral membrane proteins, offering guidelines for initial detergent selection. The differences in extraction efficiencies among this small set of detergents supports the value of detergent screening and optimization to increase the yields of targeted membrane proteins.
从脂质双分子层环境中提取后为膜蛋白创造稳定条件对于后续表征至关重要。去污剂是获得这种稳定环境最广泛使用的手段;然而,已发现不同类型的膜蛋白需要具有不同性质的去污剂,以实现最佳提取效率和提取后的稳定性。已针对从细菌和酵母中分离的膜以及一组重组靶蛋白研究了几种去污剂类型的提取情况。这些去污剂的提取效率在较高浓度下会增加,并且显示出与它们各自的临界胶束浓度(CMC)值相关。两种烷基糖去污剂,辛基-β-D-葡萄糖苷(OG)和5-环己基-1-戊基-β-D-麦芽糖苷(Cymal-5),以及一种两性离子表面活性剂,N-癸基磷酸胆碱(Fos-choline-10),通常在提取多种膜蛋白方面有效。然而,某些去污剂在提取特定类别的整合膜蛋白方面比其他去污剂更有效,为初始去污剂选择提供了指导。这一小类去污剂在提取效率上的差异支持了去污剂筛选和优化对于提高靶向膜蛋白产量的价值。