RESUMEN
We have investigated temperature dependence of Ca2+ uptake by the cardiac sarcolemmal Na(+)-Ca2+ exchanger from dog, rabbit and bullfrog. In native rabbit sarcolemmal vesicles, Ca2+ affinity of the Na(+)-Ca2+ exchanger is unchanged from 7 to 37 degrees C; however, the initial velocity of Ca2+ uptake declines much more steeply below 22 degrees C than above 22 degrees C. In native dog sarcolemma, the temperature dependence of Na(+)-Ca2+ exchange velocity is similar to that of native rabbit. However, in frog heart the velocity of Na(+)-Ca2+ exchange declines much more slowly with decreasing temperature at both temperature ranges. Reconstitution of the Na(+)-Ca2+ exchanger into artificial lipid vesicles consisting of either asolectin or phosphatidylserine, phosphatidylcholine, and cholesterol has little effect on temperature dependence of Na(+)-Ca2+ exchange velocity in any of the three species. We conclude that the lesser temperature sensitivity of the cardiac sarcolemmal Na(+)-Ca2+ exchanger of a poikilothermic species is at least partly an intrinsic property of the transport protein.
Asunto(s)
Calcio/metabolismo , Proteínas Portadoras/metabolismo , Miocardio/metabolismo , Sarcolema/metabolismo , Animales , Perros , Cinética , Conejos , Rana catesbeiana , Intercambiador de Sodio-Calcio , Especificidad de la Especie , TermodinámicaRESUMEN
Vesicles in a highly enriched sarcolemma preparation from canine ventricle were found to develop membrane potentials in response to outwardly directed potassium and inwardly directed sodium concentration gradients. The magnitude of the potential measured by the fluorescent dye diS-C3-(5) suggested a sodium-to-potassium permeability ratio between 0.2 and 1.0 which is one to two orders of magnitude greater than values obtained for the myocardial cell. Radiotracer techniques were employed to evaluate the permeability coefficients of the isolated cardiac sarcolemma membrane for sodium and potassium under equilibrium conditions (i.e., equal salt concentrations in the intravesicular and extravesicular spaces). The uptake of sodium and potassium was best described by two exponential processes which followed an increment of uptake that occurred prior to the earliest assay time (i.e., 17 sec). The compartment sizes were linear, nonsaturable functions of the cation activity. Evaluation of the rate coefficients of cation uptake by the two exponential processes versus cation activity revealed that sodium influx via the slow process and potassium influx via the fast process varied linearly with cation activity, suggesting that the permeability coefficients were concentration independent for these compartments. Conversely, sodium influx via the fast process exhibited a nonlinear increase with increasing sodium activity, and potassium influx via the slow process appeared to saturate with increasing potassium activity. In general, the permeabilities of the sarcolemma-enriched preparation for sodium and potassium were of equal magnitude. The permeability coefficients were lower than that for the potassium coefficient reported for cardiac cells but are in the range of that reported for sodium.