title: In vitro analysis of the stop-transfer process during translocation across the cytoplasmic membrane of Escherichia coli.
abstract: In this study, using a derivative of proOmpA containing an artificial stop-transfer sequence (proOmpA2xH1), we analyzed the process of stop-transfer during translocation across the cytoplasmic membrane of Escherichia coli. ProOmpA2xH1 did not interfere with the transit of wild-type proOmpA. When proOmpA2xH1 was anchored in the membrane, membrane-inserted SecA was deinserted with the reversion of the inverted topology of SecG. Cross-linking experiments revealed that the anchored proOmpA2xH1 that does not interact with either SecY or SecA. These results, taken together, suggest that proOmpA2xH1 leaves the translocation pathway by means of a specific interaction between the stop-transfer sequence and the translocational channel.
authors.affiliation: Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sato
authors.forename: H
authors.initials: H
authors.lastname: Mori
authors.forename: M
authors.initials: M
authors.lastname: Yoshida
authors.forename: M
authors.initials: M
authors.lastname: Tagaya
authors.forename: S
authors.initials: S
authors.lastname: Mizushima
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D001665:Binding Sites; D001692:Biological Transport; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006868:Hydrolysis; D007425:Intracellular Membranes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D000069816:SEC Translocation Channels
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011498:Protein Precursors; D000069816:SEC Translocation Channels; C088616:SecG protein, E coli; C057184:outer membrane protein A precursor (E coli); D000255:Adenosine Triphosphate