Regulation of K-Cl cotransport in erythrocytes of frog Rana temporaria by commonly used protein kinase and protein phosphatase inhibitors.

Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology
Gennadii Petrovich Gusev, Natalia Ivanovna Agalakova

Abstract

Recently (Agalakova and Gusev in J Comp Physiol 179:443-450, 2009), we demonstrated that the activity of K-Cl cotransport (KCC) in frog red blood cells is inhibited under stimulation of protein kinase C (PKC) with phorbol ester PMA (12-myristate-13-acetate). Present work was performed to uncover possible implication of protein kinases and protein phosphatases (PPs) in the regulation of baseline and volume-dependent KCC activity in these cells. K+ influx was estimated as 86Rb uptake by the cells in isotonic or hypotonic media in the presence of ouabain, K+ efflux was determined as the difference between K+ loss by the cells incubated in parallel in isotonic or hypotonic K(+)-free Cl(-)- and NO(3)(-)-media. Swelling of the cells in hypotonic medium was accompanied by approximately 50% activation of Cl-dependent K+ influx and efflux. Protein tyrosine kinase (PTK) inhibitor genistein (0.1 mM) stably and considerably (up to 89%) suppressed both baseline and volume-dependent KCC activity in each direction. Other PTK blockers (tyrphostin 23 and quercetin) had no influence on KCC activity in frog erythrocytes. PKC inhibitor chelerythrine (20 microM) and both PP inhibitors, fluoride (5 mM) and okadaic acid (1 microM), reduced KCC activi...Continue Reading

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Citations

Feb 4, 2011·Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology·Osvaldo CharaPablo J Schwarzbaum
Mar 10, 2017·Journal of Cellular Physiology·Rachelle Frenette-CottonPaul Isenring
Jan 8, 2014·The Biochemical Journal·Paola de Los HerosJinwei Zhang
Aug 18, 2017·American Journal of Physiology. Cell Physiology·A P GarneauP Isenring

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