Na-K-ATPase in rat cerebellar granule cells is redox sensitive.
Redox-induced regulation of the Na-K-ATPase was studied in dispersed rat cerebellar granule cells. Intracellular thiol redox state was modulated using glutathione (GSH)-conjugating agents and membrane-permeable ethyl ester of GSH (et-GSH) and Na-K-ATPase transport and hydrolytic activity monitored as a function of intracellular reduced thiol concentration. Depletion of cytosolic and mitochondrial GSH pools caused an increase in free radical production in mitochondria and rapid ATP deprivation with a subsequent decrease in transport but not hydrolytic activity of the Na-K-ATPase. Selective conjugation of cytosolic GSH did not affect free radical production and Na-K-ATPase function. Unexpectedly, overloading of cerebellar granule cells with GSH triggered global free radical burst originating most probably from GSH autooxidation. The latter was not followed by ATP depletion but resulted in suppression of active K(+) influx and a modest increase in mortality. Suppression of transport activity of the Na-K-ATPase was observed in granule cells exposed to both permeable et-GSH and impermeable GSH, with inhibitory effects of external and cytosolic GSH being additive. The obtained data indicate that redox state is a potent regulator of the Na-K-ATPase function. Shifts from an "optimal redox potential range" to higher or lower levels cause suppression of the Na-K pump activity.
Petrushanko
I
I
Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, CH-8057 Zurich, Switzerland.
Bogdanov
N
N
Bulygina
E
E
Grenacher
B
B
Leinsoo
T
T
Boldyrev
A
A
Gassmann
M
M
Bogdanova
A
A
D000255:Adenosine Triphosphate; D000818:Animals; D002470:Cell Survival; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004137:Dinitrochlorobenzene / Q000494:pharmacology*; D005260:Female; D005978:Glutathione / Q000378:metabolism* / Q000494:pharmacology; D006868:Hydrolysis; D008293:Malates / Q000494:pharmacology*; D008928:Mitochondria / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D004137:Dinitrochlorobenzene; D008293:Malates; D017382:Reactive Oxygen Species; C058705:diethyl malate; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase; D005978:Glutathione