The orientation of the major coat protein of bacteriophage f1 in the cytoplasmic membrane of Escherichia coli.
The orientation of the major coat (B) protein of the bacteriophage f1, an integral membrane protein in the cytoplasmic membrane of infected Escherichia coli, was examined. Pyridoxal 5'-phosphate and [3H]NaBH4 were used to label the cytoplasmic membrane proteins in spheroplasts and membrane vesicles of E. coli infected with bacteriophage f1. Under the conditions described, tritium incorporation was almost completely dependent on the presence of pyridoxal 5'-phosphate and little if any of the cytoplasmic proteins were labeled when the reaction was applied to intact spheroplasts. The major coat protein was isolated from the cytoplasmic membranes labeled in this manner and the chymotryptic peptides were analyzed for the presence of tritium in the pyridoxamine 5'-phosphate conjugate. When the proteins were labeled in the intact spheroplast, only the NH2-terminal chymotryptic peptide of the coat protein was labeled. If the proteins were labeled during osmotic lysis of the spheroplasts or in isolated vesicles, the chymotryptic peptide containing the COOH terminus of the coat protein as well as the NH2-terminal peptide was labeled. The NH2-terminal peptide was labeled to approximately the same extent as occurred in the intact spheroplast. These results are consistent with the hypothesis that the mature f1 coat protein asymmetrically spans the cytoplasmic membrane of the infected host with its NH2 terminus exposed on the outside and COOH terminus exposed on the cytoplasmic surface.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis*; D001894:Borohydrides; D002462:Cell Membrane / Q000032:analysis; D002918:Chymotrypsin; D003090:Coliphages / Q000032:analysis*; D004926:Escherichia coli / Q000032:analysis*; D008565:Membrane Proteins / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D011732:Pyridoxal Phosphate; D013104:Spheroplasts / Q000032:analysis; D014764:Viral Proteins / Q000032:analysis*
D001426:Bacterial Proteins; D001894:Borohydrides; D008565:Membrane Proteins; D010446:Peptide Fragments; D014764:Viral Proteins; D011732:Pyridoxal Phosphate; D002918:Chymotrypsin