UNIT 29.7 Methods for Studying Interactions of Detergents and Lipids with α-Helical and β-Barrel Integral Membrane Proteins
Published Online: 5 NOV 2013
Copyright © 2013 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Saif Hasan, S., Baniulis, D., Yamashita, E., Zhalnina, M. V., Zakharov, S. D., Stofleth, J. T. and Cramer, W. A. 2013. Methods for Studying Interactions of Detergents and Lipids with α-Helical and β-Barrel Integral Membrane Proteins. Current Protocols in Protein Science. 74:29.7:29.7.1–29.7.30.
- Published Online: 5 NOV 2013
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 b6f 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 b6f 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 b6f 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.
- vitamin B12 receptor;
- cytochrome b6f complex;
- OmpF porin