title: Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM(TM)): application to lipid-specific membrane protein topogenesis.
abstract: We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas-Houston, Medical School, Houston, TX 77030, USA.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Bogdanov
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Xie
authors.forename: William
authors.initials: W
authors.lastname: Dowhan
mesh_terms: D000465:Algorithms; D001671:Biochemistry / Q000379:methods*; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism*; D003545:Cysteine / Q000737:chemistry*; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D017930:Genes, Reporter; D006031:Glycosylation; D008051:Lipid Bilayers / Q000737:chemistry; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000378:metabolism; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008301:Maleimides / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry*; D026901:Membrane Transport Proteins / Q000737:chemistry; D008956:Models, Chemical; D009154:Mutation; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D000939:Epitopes; D008051:Lipid Bilayers; D008055:Lipids; D008081:Liposomes; D046911:Macromolecular Substances; D008301:Maleimides; D008563:Membrane Lipids; D026901:Membrane Transport Proteins; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D013438:Sulfhydryl Compounds; C483858:phosphatidylethanolamine; D003545:Cysteine