The antagonistic effect of K+o and dihydro-ouabain on the Na+ pump current of single rat and guinea-pig cardiac cells.
1. The antagonistic effect of extracellular potassium ions (K+o) and dihydro-ouabain (DHO) on the Na(+)-K+ pump current (Ip) was studied in isolated ventricular cells. 2. The myocytes were isolated from rats and guinea-pigs, two species with different sensitivity towards cardiac glycosides. Ip measurements were performed at 32-34 degrees C by means of whole-cell recording. The membrane potential was held at -20 mV throughout. 3. The DHO concentration ([DHO]) required for half-maximal Ip inhibition (apparent KD value, KD') amounted to 2.4 x 10(-3) and 1.4 x 10(-5) M for rat and guinea-pig myocytes, respectively, at 5.4 mM K+o. 4. The data suggest one-to-one binding of DHO to the Na(+)-K+ pump and a smaller association rate constant, as well as a larger dissociation rate constant, for binding of DHO in the rat cells. 5. Ip activation by K+o was nearly identical in myocytes of both species and was measured to be half-maximal at approximately 1 mM K+o. Half-maximal Ip activation by K+o remained essentially unchanged, but Ip decreased in media containing [DHO] near the respective KD' at 5.4 mM K+o. 6. The concentration-response curve of Ip inhibition by DHO was shifted to higher [DHO] at higher [K+]o. KD' increased correspondingly. The slope of the curve was unaffected. 7. Ip and KD' displayed a similar dependence on [K+]o. 8. KD' was larger in Na(+)-free than in Na(+)-containing media under conditions in which the activation of Ip by K+o was nearly the same. 9. It is concluded that the antagonism between K+o and DHO, with regard to the activation of Ip, is non-competitive. A possible mechanism of the antagonism is discussed. The mechanism implies binding of K+o and DHO to different conformational states of the Na(+)-K+ pump which are temporarily exposed to the external face of the sarcolemma in the pump cycle. The DHO-bound states do not participate in the generation of Ip.
Hermans
A N
AN
Interdisciplinary Research Centre, Catholic University of Leuven, Kortrijk, Belgium.
Glitsch
H G
HG
Verdonck
F
F
D000818:Animals; D004553:Electric Conductivity; D006168:Guinea Pigs; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010042:Ouabain / Q000031:analogs & derivatives* / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology*; D051381:Rats; D012964:Sodium / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000502:physiology*; D013045:Species Specificity
C000977:dihydroouabain; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D011188:Potassium