title: Study of polytopic membrane protein topological organization as a function of membrane lipid composition.
abstract: A protocol is described using lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by the substituted-cysteine accessibility method as applied to transmembrane domains (SCAM). SCAM is adapted to follow changes in membrane protein topology as a function of changes in membrane lipid composition. The strategy described can be adapted to any membrane system.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, TX, USA.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Bogdanov
authors.forename: Philip N
authors.initials: PN
authors.lastname: Heacock
authors.forename: William
authors.initials: W
authors.lastname: Dowhan
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D047468:Immunoprecipitation; D008563:Membrane Lipids / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000235:genetics / Q000378:metabolism; D010743:Phospholipids / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C483858:phosphatidylethanolamine; C021499:lactose permease; D016695:Glycosyltransferases; C412464:PssA protein, Rhizobium leguminosarum