Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli.
A major antigenic constituent of the inner membrane of Escherichia coli ML308-225 was identified as a 28.5-kilodalton lipoprotein containing covalently bound glycerol and palmitate. This lipoprotein corresponded to antigen 47 in the crossed immunoelectrophoresis profile of membrane vesicles (P. Owen and H.R. Kaback, Proc. Natl. Acad. Sci. USA 75:3148-3152, 1978) and to new lipoprotein 4 described for E. coli B by Ichihara et al. (S. Ichihara, H. Hussain, and S. Mizushima, J. Biol. Chem. 256:3125-3129, 1980). Experiments involving isopycnic centrifugation of spheroplast envelopes indicated that antigen 47 was enriched in cytoplasmic membrane subfractions of low density. The protein did not manifest an obvious association with peptidoglycan of the types displayed by the bound form of the Braun (Lpp) lipoprotein, the 21-kilodalton peptidoglycan-associated lipoprotein, or the ompF/C gene products. Antibodies specific for antigen 47 were used to demonstrate that the molecule was immunologically distinct from both the Braun lipoprotein and the peptidoglycan-associated lipoprotein of E. coli. Antigens of similar molecular mass to and cross-reacting with antigen 47 were present in the envelopes of eight type species of the Enterobacteriaceae. A protocol for the purification of antigen 47, based upon its solubility in a chloroform-methanol-water mixture, was developed.
D000942:Antigens, Bacterial / Q000032:analysis*; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis*; D007123:Immunoelectrophoresis, Two-Dimensional; D007425:Intracellular Membranes / Q000032:analysis* / Q000276:immunology; D008074:Lipoproteins / Q000032:analysis / Q000276:immunology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D017830:Octoxynol; D010457:Peptidoglycan / Q000378:metabolism; D011092:Polyethylene Glycols
D000942:Antigens, Bacterial; D008074:Lipoproteins; D046911:Macromolecular Substances; D010457:Peptidoglycan; D011092:Polyethylene Glycols; D017830:Octoxynol