Document Type

Article

Publication Date

1995

Abstract

45Ca2+ uptake by purified brush-border membrane vesicles of starfish (Pycnopodia helianthoides) pyloric ceca was stimulated by an outwardly directed H+ gradient and this stimulation was enhanced by the simultaneous presence of an induced membrane potential (inside negative; K+/valinomycin). External amiloride (competitive inhibitor; Ki=660mmol l21) and a monoclonal antibody raised against proteins associated with the lobster (Homarus americanus) electrogenic 2Na+/1H+ antiporter both inhibited approximately half of the proton-gradient stimulated 45Ca2+ uptake. These results suggested that Ca2+ might be transported by the electrogenic antiporter and that the crustacean antibody was inhibitory to the exchange function in echinoderms, as was recently shown in crustacean epithelial brush-border membrane vesicles. Carrier-mediated 45Ca2+ influx by amiloride-sensitive and amiloride-insensitive systems displayed the following kinetic constants: (amiloride-sensitive) Kt=66±2 mmol l21; Jmax=0.173±0.002 pmol mg21 protein 8 s21; (amiloride-insensitive) Kt=18±0.3 mmol l21; Jmax=0.100±0.001 pmol mg21 protein 8 s21. Zn2+ was a mixed inhibitor of 45Ca2+ influx by carrier-mediated transport, displaying a Ki of 920mmol l21. Mn2+, Cu2+, Fe2+ and Mg2+ also inhibited 45Ca2+ uptake, but the mechanism(s) of inhibition by these other cations was not disclosed. An equilibrium shift experiment showed that both Na+ and Zn2+ were able to exchange with equilibrated 45Ca2+ in these vesicles, suggesting that both monovalent and divalent cations were able to enter pyloric cecal cells through a common carrier-mediated transport system. In addition, the echinoderm electrogenic system appeared to exhibit a molecular component recognized by the crustacean antibody that may imply a similar epitope in the two animals.

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