Unit

UNIT 29.7 Methods for Studying Interactions of Detergents and Lipids with α-Helical and β-Barrel Integral Membrane Proteins

  1. S. Saif Hasan1,
  2. Danas Baniulis1,2,
  3. Eiki Yamashita3,
  4. Mariya V. Zhalnina1,
  5. Stanislav D. Zakharov1,4,
  6. Jason T. Stofleth1,5,
  7. William A. Cramer1

Published Online: 5 NOV 2013

DOI: 10.1002/0471140864.ps2907s74

Current Protocols in Protein Science

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.

Author Information

  1. 1

    Department of Biological Sciences, Purdue University, West Lafayette, Indiana

  2. 2

    Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, Babtai, Kaunas Region, Lithuania

  3. 3

    Institute for Protein Research, Osaka University, Suita, Osaka, Japan

  4. 4

    Institute of Basic Problems of Biology, Russian Academy of Sciences, Pushchino, Moscow Region, Russian Federation

  5. 5

    Department of Chemistry and Biochemistry, University of California San Diego, San Diego, California

Publication History

  1. Published Online: 5 NOV 2013

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 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.

Keywords:

  • BtuB;
  • vitamin B12 receptor;
  • cytochrome b6f complex;
  • OmpF porin