Methods for Studying Interactions of Detergents and Lipids with α‐Helical and β‐Barrel Integral Membrane Proteins. Issue 1 (16th February 2018)

• Record Type: Journal Article
• Authors: Saif Hasan, S.; Baniulis, Danas; Yamashita, Eiki; Zhalnina, Mariya V.; Zakharov, Stanislav D.; Stofleth, Jason T.; Cramer, William A.
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 74:Issue 1(2013)
• Page Start: 29.7.1
• Page End: 29.7.30
• Publication Date: 2018-02-16
• Subjects: BtuB -- vitamin B12 receptor -- cytochrome b6f complex -- OmpF porin; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/0471140864.ps2907s74
• Languages: English
• ISSNs: 1934-3655

Abstract

Abstract: Methods for studying interactions of protein with lipids and detergents are described for representatives of two major classes of membrane proteins: (1) the α‐helical hetero‐oligomeric integral cytochrome b 6 f complex of oxygenic photosynthesis from cyanobacteria, and (2) the outer membrane β‐barrel proteins BtuB and OmpF from Gram‐negative Escherichia coli bacteria. Details are presented on the use of detergents for purification and crystallization of the b 6 f complex as well as a method for lipid exchange. The positions of detergent and lipid molecules, which define eight potential lipid‐binding sites in the b 6 f complex, are described. Differences in detergent strategies for isolation and crystallization of β‐barrel proteins relative to those for oligomeric helical membrane proteins are discussed, and purification and assessment of protein quality by circular dichroism (CD) is presented. Curr. Protoc. Protein Sci . 74:29.7.1‐29.7.30. © 2013 by John Wiley & Sons, Inc.