Use of a foreign epitope as a "tag" for the localization of minor proteins within a cell: the case of the immunity protein to colicin A.
The immunity protein to colicin A (Cai), which is constitutively expressed at a very low level in Escherichia coli strains, has been studied in recombinant plasmid constructs allowing expression of various immunity fusion proteins under the control of inducible promoters. The 13-amino acid NH2-terminal region of Cai was substituted by polypeptides from beta-galactosidase or from colicin A. Upon induction of the chimeric proteins, the rate of expression of the immunity protein could be correlated to the level of resistance to colicin A. The immunity protein has been "tagged" with an epitope from the colicin A protein for which a monoclonal antibody is available. Using this technique, we have directly demonstrated that the immunity protein is located in the cytoplasmic membrane. The results indicate that the NH2-terminal region of Cai is directed toward the cytoplasm and is probably not required for Cai insertion into the membrane or for its function.
D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D003087:Colicins / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000032:analysis* / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis*; D001616:beta-Galactosidase / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D003087:Colicins; D000939:Epitopes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C007104:colicin immunity proteins; D001616:beta-Galactosidase