title: Methods for studying interactions of detergents and lipids with α-helical and β-barrel integral membrane proteins.
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.
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
authors: S
authors: S
authors: Saif Hasan
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.|Institute of Horticulture, Lithuanian Research Centre for Agriculture and Forestry, Babtai, Kaunas Region, Lithuania.
authors: Danas
authors: D
authors: Baniulis
authors: Institute for Protein Research, Osaka University, Suita, Osaka, Japan.
authors: Eiki
authors: E
authors: Yamashita
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
authors: Mariya V
authors: MV
authors: Zhalnina
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.|Institute of Basic Problems of Biology, Russian Academy of Sciences, Pushchino, Moscow Region, Russian Federation.
authors: Stanislav D
authors: SD
authors: Zakharov
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.|Department of Chemistry and Biochemistry, University of California San Diego, San Diego, California.
authors: Jason T
authors: JT
authors: Stofleth
authors: Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
authors: William A
authors: WA
authors: Cramer
mesh_terms: D001425:Bacterial Outer Membrane Proteins; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D003460:Crystallization; D045346:Cytochrome b6f Complex; D003902:Detergents / Q000737:chemistry*; D029968:Escherichia coli Proteins; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D026901:Membrane Transport Proteins; D008958:Models, Molecular; D018272:Porins; D017433:Protein Structure, Secondary
chemical_list: D001425:Bacterial Outer Membrane Proteins; C044592:BtuB protein, E coli; D003902:Detergents; D029968:Escherichia coli Proteins; D008055:Lipids; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C109237:OmpF protein; D018272:Porins; D045346:Cytochrome b6f Complex
keywords: BtuB; OmpF porin; cytochrome b6f complex; vitamin B12 receptor