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Title: Proton-stimulated Cl-HCO/sub 3/ antiport by basolateral membrane vesicles of lobster hepatopancreas

Journal Article · · Am. J. Physiol.; (United States)
OSTI ID:6409361

Purified epithelial basolateral membrane vesicles were prepared from lobster hepatopancreas by sorbitol gradient centrifugation. Na+-K+-adenosinetriphosphatase, alkaline phosphatase, and cytochrome-c oxidase enzyme activities in the final membrane preparation were enriched 9.6-, 1.4-, and 0.4-fold, respectively, compared with their activities in the original tissue homogenate. Vesicle osmotic reactivity was demonstrated using 60-min equilibrium /sup 36/Cl uptake experiments at a variety of transmembrane osmotic gradients. /sup 36/Cl uptake into vesicles preloaded with HCO/sub 3/ was significantly greater than into vesicles lacking HCO/sub 3/. This exchange process was stimulated by a transmembrane proton gradient (internal pH greater than external pH). Proton-gradient-dependent Cl-HCO/sub 3/ exchange was potential sensitive and stimulated by an electrically negative vesicle interior. /sup 36/Cl influx (4-s exposures) into HCO/sub 3/-loaded vesicles occurred by the combination of 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid sensitive, carrier-mediated transfer and apparent diffusion. /sup 36/Cl influx was a hyperbolic function of both internal (HCO/sub 3/) and internal (Cl). The two internal anions displayed a 100-fold difference in apparent affinity constants with HCO/sub 3/ being strongly preferred. /sup 36/Cl influx was stimulated more by preloaded monovalent than by divalent anions. Na was an inhibitor of proton-dependent anion antiport, whereas K had no effect. A model for HCl-HCO/sub 3/ antiport is suggested that employs combined transmembrane concentration gradients of Cl and HCO/sub 3/ to power anion exchange and transfer protons against a concentration gradient.

Research Organization:
Univ. of Hawaii, Manoa, Honolulu
OSTI ID:
6409361
Journal Information:
Am. J. Physiol.; (United States), Journal Name: Am. J. Physiol.; (United States)
Country of Publication:
United States
Language:
English

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