title: Assembly of bacterial outer membrane proteins.
abstract: Various methods that are routinely used to study the subcellular localization of membrane proteins in wild-type Gram-negative bacteria fall short in genetic studies addressing the biogenesis of outer membrane proteins (OMPs). Here, we describe three biochemical methods that can be used in such studies to evaluate the proper assembly of OMPs into the outer membrane. The methods are based on (1) the differential electrophoretic mobility of folded and nonnative OMPs, (2) the intrinsically high protease resistance of folded OMPs, and (3) the observation that integral membrane proteins are not extracted from the membrane in solutions containing high concentrations of urea.
authors.affiliation: Department of Molecular Microbiology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Grijpstra
authors.forename: Martine P
authors.initials: MP
authors.lastname: Bos
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay
chemical_list: D001425:Bacterial Outer Membrane Proteins