Effect of taurine on chelerythrine inhibition of calcium uptake and ATPase activity in the rat retina.
Taurine potentiates calcium uptake in whole retinal homogenates as well as in rod outer segments and mitochondrial fractions. The aim of this study was to correlate taurine potentiation of calcium uptake with its effects on other cellular processes through the use of chelerythrine (CHT), a modulator of protein kinase C (PKC), ATPase activity, and, as recently shown, of retinal protein phosphorylation. CHT inhibited calcium uptake only when ATP was present, and inhibition increased significantly in conditions of ATP excess. Taurine potentiated ATP-dependent calcium uptake but decreased the potency of ATP to induce uptake activity. CHT inhibition of calcium uptake exhibited similar potencies in the presence and absence of taurine, and this inhibition seemed to be independent of PKC inhibition. Because of the ATP-dependence of the observed effect, total ATPase activity was studied using similar treatments. In the absence of taurine, CHT inhibited ATPase activity with the same potency (IC50 approximately 59.3 microM) as with calcium uptake inhibition (IC50 approximately 87.9 microM), presenting a possible mechanism of action of CHT. In the presence of taurine, no such correlation was observed, suggesting an ATPase-independent mechanism of action. In fact, taurine did not potentiate ATPase activity, but rather it decreased the potency of CHT inhibition of ATPase, effects incongruent with the effects of taurine on calcium uptake and on CHT inhibition of calcium uptake. Enzyme kinetic experiments provided more supporting data. Taurine was found to cause an increase in the affinity of the ATP substrate for the ATPase enzyme, contradicting the aforementioned effect of taurine to decrease the potency of ATP to induce calcium uptake. Thus, taurine seems to increase calcium uptake through a hitherto unreported mechanism distinct from its modulation of ATPase activity.
Militante
J D
JD
Department of Pharmacology, Texas Tech University Health Sciences Center, Lubbock 79430, USA.
Lombardini
J B
JB
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002118:Calcium / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007700:Kinetics; D010617:Phenanthridines / Q000494:pharmacology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D013654:Taurine / Q000494:pharmacology*
D000470:Alkaloids; D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; D010617:Phenanthridines; D013654:Taurine; C016299:chelerythrine; D011493:Protein Kinase C; D000251:Adenosine Triphosphatases; D002118:Calcium