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Evidence from simultaneous intracellular- and surface-pH transients that carbonic anhydrase II enhances CO2 fluxes across Xenopus oocyte plasma membranes.
Musa-Aziz, Raif; Occhipinti, Rossana; Boron, Walter F.
Afiliação
  • Musa-Aziz R; Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio; Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut; and Department of Physiology and Biophysics, University of Sao Paulo, Institute of Biomedical Sciences, Sao Paulo, Brazil raifaziz@icb.usp.br.
  • Occhipinti R; Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio;
  • Boron WF; Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio; Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio; Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut; and.
Am J Physiol Cell Physiol ; 307(9): C791-813, 2014 Nov 01.
Article em En | MEDLINE | ID: mdl-24965587
The α-carbonic anhydrases (CAs) are zinc-containing enzymes that catalyze the interconversion of CO2 and HCO3 (-). Here, we focus on human CA II (CA II), a ubiquitous cytoplasmic enzyme. In the second paper in this series, we examine CA IV at the extracellular surface. After microinjecting recombinant CA II in a Tris solution (or just Tris) into oocytes, we expose oocytes to 1.5% CO2/10 mM HCO3 (-)/pH 7.50 while using microelectrodes to monitor intracellular pH (pHi) and surface pH (pHS). CO2 influx causes the familiar sustained pHi fall as well as a transient pHS rise; CO2 efflux does the opposite. Both during CO2 addition and removal, CA II increases the magnitudes of the maximal rate of pHi change, (dpHi/dt)max, and the maximal change in pHS, ΔpHS. Preincubating oocytes with the inhibitor ethoxzolamide eliminates the effects of CA II. Compared with pHS, pHi begins to change only after a delay of ~9 s and its relaxation has a larger (i.e., slower) time constant (τpHi > τpHS ). Simultaneous measurements with two pHi electrodes, one superficial and one deep, suggest that impalement depth contributes to pHi delay and higher τpHi . Using higher CO2/HCO3 (-) levels, i.e., 5%/33 mM HCO3 (-) or 10%/66 mM HCO3 (-), increases (dpHi/dt)max and ΔpHS, though not in proportion to the increase in [CO2]. A reaction-diffusion mathematical model (described in the third paper in this series) accounts for the above general features and supports the conclusion that cytosolic CA-consuming entering CO2 or replenishing exiting CO2-increases CO2 fluxes across the cell membrane.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Dióxido de Carbono / Membrana Celular / Anidrase Carbônica II Limite: Animals / Humans Idioma: En Revista: Am J Physiol Cell Physiol Assunto da revista: FISIOLOGIA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Dióxido de Carbono / Membrana Celular / Anidrase Carbônica II Limite: Animals / Humans Idioma: En Revista: Am J Physiol Cell Physiol Assunto da revista: FISIOLOGIA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos