TY - JOUR
T1 - Patch clamp studies on ion pumps of the cytoplasmic membrane of Escherichia coli
T2 - Formation, preparation, and utilization of giant vacuole- like structures consisting of everted cytoplasmic membrane
AU - Kuroda, Teruo
AU - Okuda, Naoyuki
AU - Saitoh, Naoto
AU - Hiyama, Tetsuo
AU - Terasaki, Yoko
AU - Anazawa, Hideharu
AU - Hirata, Aiko
AU - Mogi, Tatsushi
AU - Kusaka, Iwao
AU - Tsuchiya, Tomofusa
AU - Yabe, Isamu
PY - 1998/7/3
Y1 - 1998/7/3
N2 - 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 F(o)F1-ATPase. Such current was inhibited by inhibitors of the respiratory chain or F(o)F1-ATPase. This method is applicable for analyses of ion channels, ion pumps, or ion transporters in E. coli or other microorganisms.
AB - 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 F(o)F1-ATPase. Such current was inhibited by inhibitors of the respiratory chain or F(o)F1-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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U2 - 10.1074/jbc.273.27.16897
DO - 10.1074/jbc.273.27.16897
M3 - Article
C2 - 9642251
AN - SCOPUS:15644371371
VL - 273
SP - 16897
EP - 16904
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 27
ER -