title: Patch clamp studies on ion pumps of the cytoplasmic membrane of Escherichia coli. Formation, preparation, and utilization of giant vacuole-like structures consisting of everted cytoplasmic membrane.
abstract: Formation of giant protoplasts from normal Escherichia coli cells resulted in the formation of giant vacuole-type structures (which we designate as provacuoles) in the protoplasts. Electron microscopic observation revealed that these provacuoles were surrounded by a single membrane. We detected inner (cytoplasmic) membrane proteins in the provacuolar membrane but not outer membrane proteins. Biochemical analyses revealed that the provacuoles consist of everted cytoplasmic membranes. We applied the patch clamp method to the giant provacuoles. We have succeeded in measuring current that represents inward movement of H+ because of respiration and to ATP hydrolysis by the FoF1-ATPase. Such current was inhibited by inhibitors of the respiratory chain or FoF1-ATPase. This method is applicable for analyses of ion channels, ion pumps, or ion transporters in E. coli or other microorganisms.
authors.affiliation: Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama, 700, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kuroda
authors.forename: N
authors.initials: N
authors.lastname: Okuda
authors.forename: N
authors.initials: N
authors.lastname: Saitoh
authors.forename: T
authors.initials: T
authors.lastname: Hiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Terasaki
authors.forename: H
authors.initials: H
authors.lastname: Anazawa
authors.forename: A
authors.initials: A
authors.lastname: Hirata
authors.forename: T
authors.initials: T
authors.lastname: Mogi
authors.forename: I
authors.initials: I
authors.lastname: Kusaka
authors.forename: T
authors.initials: T
authors.lastname: Tsuchiya
authors.forename: I
authors.initials: I
authors.lastname: Yabe
mesh_terms: D003593:Cytoplasm / Q000378:metabolism*; D004579:Electron Transport; D004926:Escherichia coli / Q000378:metabolism* / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000378:metabolism; D016623:Ion Pumps / Q000378:metabolism* / Q000502:physiology; D008854:Microscopy, Electron; D018408:Patch-Clamp Techniques; D006180:Proton-Translocating ATPases / Q000378:metabolism
chemical_list: D016623:Ion Pumps; D006180:Proton-Translocating ATPases