Immunochemical analysis of membrane vesicles from Escherichia coli.
Membrane vesicles isolated from Escherichia coli ML 308--225 have been analyzed by crossed immunoelectrophoresis, and immunoprecipitates corresponding to the following cellular components have been identified: ATPase (EC 3.6.1,3), two or three NADH dehydrogenases (EC 1.6.99.3), D-lactate dehydrogenase (EC 1.1.1.27), glutamate dehydrogenase (EC 1.4.1.4), dihydro-orotate dehydrogenase (EC 1.3.3.1), 6-phosphogluconate dehydrogenase (EC 1.1.1.43), polynucleotide phosphorylase (EC 2.3.7.8), beta-galactosidase (EC 3.2.1.23), lipopolysaccharide, and Braun's lipoprotein. The cellular origin of many of the vesicle immunogens is determined, and Braun's lipoprotein is used as a marker to quantitate the extent of outer membrane contamination (less than 3%). Membrane antigens are also characterized with regard to their amphiphilic or hydrophilic properties by charge-shift crossed immunoelectrophoresis. Furthermore, the following immunogens cross-react with components in membrane vesicles prepared from Salmonella typhimurium: one of the three NADH dehydrogenases, ATPase, polynucleotide phosphorylase, 6-phosphogluconate dehydrogenase, Braun's lipoprotein, and three unidentified antigens. In the accompanying paper [Owen, P., & Kaback, H. R. (1979) Biochemistry 18 (following paper in this issue)] quantitative immunoadsorption is utilized to establish the topology of the vesicles with respect to the distribution of antigens on the inner and outer faces of the membrane.
D000251:Adenosine Triphosphatases / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis*; D002462:Cell Membrane / Q000276:immunology*; D004081:Dihydroorotate Oxidase / Q000032:analysis; D004926:Escherichia coli / Q000276:immunology*; D005969:Glutamate Dehydrogenase / Q000032:analysis; D007123:Immunoelectrophoresis, Two-Dimensional; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008070:Lipopolysaccharides / Q000032:analysis; D008074:Lipoproteins / Q000032:analysis; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis; D010734:Phosphogluconate Dehydrogenase / Q000032:analysis; D011117:Polyribonucleotide Nucleotidyltransferase / Q000032:analysis; D011233:Precipitin Tests; D001616:beta-Galactosidase / Q000032:analysis
D000942:Antigens, Bacterial; D008070:Lipopolysaccharides; D008074:Lipoproteins; D007770:L-Lactate Dehydrogenase; D010734:Phosphogluconate Dehydrogenase; D004081:Dihydroorotate Oxidase; D005969:Glutamate Dehydrogenase; D009247:NADH, NADPH Oxidoreductases; D011117:Polyribonucleotide Nucleotidyltransferase; D001616:beta-Galactosidase; D000251:Adenosine Triphosphatases