Methods for studying interactions of detergents and lipids with α-helical and β-barrel integral membrane proteins.

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TitleMethods for studying interactions of detergents and lipids with α-helical and β-barrel integral membrane proteins.
Publication TypeJournal Article
Year of Publication2013
AuthorsS Hasan, S, Baniulis, D, Yamashita, E, Zhalnina, MV, Zakharov, SD, Stofleth, JT, Cramer, WA
JournalCurr Protoc Protein Sci
Volume74
Pagination29.7.1-29.7.30
Date Published2013 Nov 05
ISSN1934-3663
KeywordsBacterial Outer Membrane Proteins, Chromatography, Affinity, Circular Dichroism, Crystallization, Cytochrome b6f Complex, Detergents, Escherichia coli Proteins, Lipids, Membrane Proteins, Membrane Transport Proteins, Models, Molecular, Porins, Protein Structure, Secondary
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 b6 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 b6 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 b6 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.

DOI10.1002/0471140864.ps2907s74
Alternate JournalCurr Protoc Protein Sci
PubMed ID24510648
PubMed Central IDPMC4062877
Grant ListR01 GM018457 / GM / NIGMS NIH HHS / United States
R01 GM038323 / GM / NIGMS NIH HHS / United States
GM-018457 / GM / NIGMS NIH HHS / United States
GM-038323 / GM / NIGMS NIH HHS / United States