title: A high concentration of SecA allows proton motive force-independent translocation of a model secretory protein into Escherichia coli membrane vesicles.
abstract: The in vitro translocation of OmpF-Lpp, a model secretory protein, into inverted membrane vesicles of Escherichia coli obligatorily requires the proton motive force (delta mu H+) in the conventional assay system (Yamada, H., Tokuda, H., and Mizushima, S. (1989) J. Biol. Chem. 264, 1723-1728). The translocation, however, took place efficiently, even in the absence of delta mu H+, when the system was supplemented with additional SecA. With the stripped membrane vesicles, which are permeable to protons, or in the absence of NADH, the supplementation of SecA remarkably stimulated the translocation activity. The further addition of NADH did not significantly enhance the translocation activity under the SecA-enriched conditions. OmpF-Lpp thus translocated could be recovered from the vesicular lumen by sonication, indicating that complete translocation occurred in the absence of delta mu H+. It is suggested that delta mu H+ is required for high affinity interaction of SecA with the presumed secretory machinery in the cytoplasmic membrane and that a high concentration of SecA modulates the delta mu H+ requirement.
authors.affiliation: Laboratory of Microbiology, School of Agriculture, Nagoya University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yamada
authors.forename: S
authors.initials: S
authors.lastname: Matsuyama
authors.forename: H
authors.initials: H
authors.lastname: Tokuda
authors.forename: S
authors.initials: S
authors.lastname: Mizushima
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000494:pharmacology*; D001693:Biological Transport, Active / Q000187:drug effects; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D004563:Electrochemistry; D004926:Escherichia coli / Q000378:metabolism*; D008564:Membrane Potentials; D009243:NAD / Q000494:pharmacology; D011522:Protons
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D011522:Protons; D009243:NAD; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000255:Adenosine Triphosphate