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  • Title: The cardiac Na(+)-Ca2+ exchanger: relative rates of calcium and sodium movements and their modulation by protonation-deprotonation of the carrier.
    Author: Khananshvili D, Weil-Maslansky E.
    Journal: Biochemistry; 1994 Jan 11; 33(1):312-9. PubMed ID: 8286352.
    Abstract:
    The exchange cycle of the cardiac Na(+)-Ca2+ exchanger can be described as separate steps of Ca2+ and Na+ transport [Khananshvili, D. (1990) Biochemistry 29, 2437-2442]. In order to determine the relative rates of Na+ and Ca2+ movement during the Na(+)-Ca2+ and Ca(2+)-Ca2+ exchange modes, the ratios (R) of Na(+)-Ca2+/Ca(2+)-Ca2+ exchanges were estimated with saturating concentrations of ions at both sides of the membrane. The effect of extravesicular pH and voltage (potassium valinomycin) on the initial rates (t = 1 s) of Na(+)-Ca2+ and Ca(2+)-Ca2+ exchange were investigated by assuming that, under the conditions tested, the intravesicular pH (pH 7.4) is not affected. Na(+)- or Ca(2+)-preloaded sarcolemma vesicles were diluted rapidly in assay medium containing 45Ca and buffer (pH 5.0-10.9), and the reaction of 45Ca uptake was quenched by using a semi-rapid-mixing device. Under conditions in which [45Ca]o = [Ca]i = 250 microM, the pH-dependent curve of Ca(2+)-Ca2+ exchange shows a bell shape in the acidic range (pKa1 = 5.1 +/- 0.1 and pKa2 = 6.5 +/- 0.2) followed by activation of the exchange in the alkaline range (pKa3 = 10.0 +/- 0.2). With [45Ca]o = 250 microM and [Na]i = 160 mM, the Na(+)-Ca2+ exchange increases monotonically from pH 5.0 to 9.5 (pKa1 = 5.1 +/- 0.1, pKa2 = 7.2 +/- 0.2, and pKa3 = 9.1 +/- 0.2). At pH < 6.1, the ratio of Na(+)-Ca2+/Ca(2+)-Ca2+ exchange is close to unity (R approximately 1), while it increases to R = 3-4 in the range of pH 7.1-9.3.(ABSTRACT TRUNCATED AT 250 WORDS)
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