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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Nov;87(21):8575–8579. doi: 10.1073/pnas.87.21.8575

Volume-sensitive K-Cl cotransport in inside-out vesicles made from erythrocyte membranes from sheep of low-K phenotype.

G R Kracke 1, P B Dunham 1
PMCID: PMC54999  PMID: 2236068

Abstract

Unidirectional K ion effluxes were measured from inside-out vesicles prepared from erythrocyte membranes from sheep of the low-K phenotype. Total K efflux was 150 nmol per mg of protein per hr in a Cl medium of 295 mosmol/kg (with the Na/K pump inhibited). Cl-dependent K efflux (determined with methanesulfonate replacing Cl) was 54 nmol/(mg.hr). Cl-dependent K efflux (K-Cl cotransport) increased to 77 nmol/(mg.hr) with osmotic swelling of approximately 30% in 230-mosmol/kg medium and decreased to 13 nmol/(mg.hr) after shrinkage of approximately 60% in 430-mosmol/kg medium. Osmotically induced changes in transport and vesicle volume were reversible. K-Cl cotransport was enhanced by ATP. Nonhydrolyzable ATP analogues failed to substitute for ATP, indicating that phosphorylation is involved. However, in the absence of added ATP there was significant K-Cl cotransport, suggesting that phosphorylation is not essential for function. The results provide clues about the nature of the signals detected by the sensor of cell volume changes and demonstrate that inside-out vesicles from sheep erythrocyte membranes provide an advantageous experimental system for investigation of the volume sensor.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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