Azide- and Vanadate-Sensitive M-Phase Alkalinity and Cytosolic Acidification of Chang Liver Cells.

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概要

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• Flow cytometric cell-by-cell evaluation of NH<SUB>4</SUB>Cl acidification of human Chang cells showed that at steady state, 3% of the cells remained alkalinized (>pH<SUB>i</SUB> 7) over an extended period (up to 80 min) despite the absence of extracellular Na<SUP>+</SUP> and HCO<SUB>3</SUB><SUP>-</SUP>. In fluorescence microscopy, the acidification-resistant cells were characteristically rounded M-phase cells. Both mean cytosolic pH and M-phase alkalinity were however sensitive to (a) azide and oligomycin, inhibitors of F-ATPase (ATP synthase), and to (b) vanadium ions, the phosphate analogue of P-ATPase (ATP-hydrolyzing), in dose-dependent and time-dependent man ners. Dead cell indices were constant at ≈ 10%. Thiocyanate chaotrophic anions, which cleave the V-ATPase structure, had no effect. Since ATP synthesizing F-ATPase (ATP synthase) is coupled to ATP-hydrolyzing P-ATPase as ''master-&-slave'', azide- and oligomycin-sensitivity corroborated with vanadate-sensitivity in suggesting energized proton pumping modulating (a) M-phase alkalinity and (b) cytosolic pH, against acidification.

• 公益社団法人 日本薬理学会の論文

著者

• Bay Boon-huat

  Department Of Anatomy National University Of Singapore

• Paramanantham Ramanujam

  Department of Anatomy, Faculty of Medicine, National University of Singapore

• Wong Kim-Ping

  Department of Biochemistry, Faculty of Medicine, National University of Singapore

• Sit Kwok-Hung

  Department of Anatomy, Faculty of Medicine, National University of Singapore

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