Peptide-specific antibody locates the COOH terminus of the lactose carrier of Escherichia coli on the cytoplasmic side of the plasma membrane.
The carboxyl-terminal decapeptide NH2-Leu-Leu-Arg-Arg-Gln-Val-Asn-Glu-Val-Ala-OH of the lactose carrier protein, the product of the lac Y gene of Escherichia coli, was synthesized, and specific anti-peptide antibodies were raised in rabbits. These antibodies bind to membrane-bound lactose carrier showing that the carboxyl terminus is accessible from the aqueous phase. The antibodies bind only to the surface of inverted cytoplasmic membrane vesicles (but not to closed, right-side-out membrane vesicles), demonstrating that the carboxyl terminus of the carrier protein is directed towards the cytoplasmic side of the plasma membrane in cells. The carboxyl terminus is a potent immunogenic epitope on the purified, detergent-solubilized carrier. Binding of peptide-specific antibodies to the carrier protein inhibits neither substrate binding nor translocation.
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D002462:Cell Membrane / Q000032:analysis; D003593:Cytoplasm; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D026901:Membrane Transport Proteins / Q000276:immunology*; D009004:Monosaccharide Transport Proteins; D027981:Symporters
D029968:Escherichia coli Proteins; D007140:Immunoglobulin Fab Fragments; C050221:LacY protein, E coli; D026901:Membrane Transport Proteins; D009004:Monosaccharide Transport Proteins; D027981:Symporters; C021499:lactose permease