Orientation of the carboxyl terminus of the Na+/proline symport carrier in Escherichia coli.
The orientation of the carboxyl terminal region of the Escherichia coli proline carrier in the cytoplasmic membrane was studied. The beta-galactosidase moiety of the PutP-LacZ fusion protein [(1987) J. Biol. Chem. 262, 14100-14104] was exposed outside the inside-out vesicles and inside the right-side-out vesicles. A site-directed antibody raised against a synthetic peptide corresponding to the putative carboxyl terminal region of the carrier reacted preferentially with the inside-out vesicles prepared from a wild-type proline carrier overproducing strain and less with the right-side-out vesicles. These results indicate that the carboxyl terminus of the proline carrier is exposed to the cytoplasmic side of the membrane.
Komeiji
Y
Y
Department of Biology, Faculty of Science, University of Tokyo, Japan.
Hanada
K
K
Yamato
I
I
Anraku
Y
Y
D026921:Amino Acid Transport Systems, Neutral; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005696:Galactosidases / Q000378:metabolism*; D007763:Lac Operon; D015394:Molecular Structure; D011392:Proline / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D012964:Sodium / Q000378:metabolism*; D027981:Symporters; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
D026921:Amino Acid Transport Systems, Neutral; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D027981:Symporters; C041832:proline transport protein; C060609:PutP protein, E coli; D011392:Proline; D012964:Sodium; D005696:Galactosidases; D001616:beta-Galactosidase