Immunochemical analysis of membrane vesicles from Escherichia coli

Owen, P.; Kaback, H.R.

Biochemistry 18(8): 1413-1422


ISSN/ISBN: 0006-2960
PMID: 218620
DOI: 10.1021/bi00575a004
Accession: 040356715

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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, D-lactate dehydrogenase (EC, glutamate dehydrogenase (EC, dihydro-orotate dehydrogenase (EC, 6-phosphogluconate dehydrogenase (EC, polynucleotide phosphorylase (EC, beta-galactosidase (EC, 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.