Characterising protein/detergent complexes by triple-detection size-exclusion chromatography

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• 作者：Katharina Gimpl ; Jessica Klement ; Sandro Keller
• 关键词：Membrane proteins ; Detergent micelles ; Absolute mass determination ; Multiple detection ; Static light scattering
• 刊名：Biological Procedures Online
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：18
• 期：1
• 全文大小：1,195 KB
• 刊物主题：Biomedicine general; Biological Techniques;
• 出版者：BioMed Central
• ISSN：1480-9222

文摘

Background In vitro investigations of membrane proteins usually depend on detergents for protein solubilisation and stabilisation. The amount of detergent bound to a membrane protein is relevant to successful experiment design and data analysis but is often unknown. Triple-detection size-exclusion chromatography enables simultaneous separation of protein/detergent complexes and protein-free detergent micelles and determination of their molar masses in a straightforward and absolute manner. Size-exclusion chromatography is used to separate different species, while ultraviolet absorbance, static light scattering, and refractive index measurements allow molar mass determination of protein and detergent components.