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.

The Journal of Biological Chemistry
T KurodaIsamu Yabe

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.

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Citations

Aug 21, 2002·Biological & Pharmaceutical Bulletin·Tadazumi KomiyamaYasuhiro Furuichi
Dec 15, 2010·Bioscience, Biotechnology, and Biochemistry·Ken-ichi HashimotoHisashi Kawasaki
Sep 15, 2011·The Journal of General and Applied Microbiology·Taihei FukudomiIsamu Miyakawa
Jul 13, 2012·Bioscience, Biotechnology, and Biochemistry·Ken-ichi HashimotoHisashi Kawasaki
Jan 20, 2016·The Journal of General and Applied Microbiology·Sawako Takahashi, Hiromi Nishida
Apr 28, 2009·Journal of Bioscience and Bioengineering·Ruei-Yu Li, Chu-Yuan Cheng
May 29, 2015·The Journal of General and Applied Microbiology·Akane NojiriHiromi Nishida
Jan 25, 2011·Biochimica Et Biophysica Acta·Koji NakamuraIsamu Yabe
Sep 18, 2018·Microbiology·Koki NishinoHiromi Nishida
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