Ion channel-like properties of the Na+/K+ Pump.
Ion pumps and exchangers are considered to be different from ion channels for two principal reasons. Ion pumps move ions against, whereas ion channels allow ions to move with, the electrochemical potential gradient, and pumps transport ions relatively slowly, approximately 10(2) s(-1), whereas channels conduct ions rapidly, approximately 10(7) s(-1). However, the latter high rate refers only to the open pore, and yet all ion channels contain at least one gate. Not surprisingly, the conformational changes associated with channel gating occur with kinetics similar to those of ion pumping. Indeed, ion pumps may be viewed as ion channels with two gates, one external to, and the other internal to, the ion binding cavity. The simple operational rule for such a pump is that the two gates should never be open simultaneously; otherwise, the pump would become a channel and conduct dissipative fluxes several orders of magnitude larger than, and in the opposite direction to, the active transport fluxes. Analyses of Na(+) ion movements mediated by the Na(+)/K(+) pump under various conditions have suggested that in at least one, short-lived, conformation of the pump, an ion-channel-like structure, closed at its intracellular end, connects the extracellular solution with the ion binding sites deep in the protein core. Here we use the marine toxin, palytoxin, to act on Na(+)/K(+) pumps in outside-out patches excised from cardiac myocytes and so transform the pumps into nonselective cation channels which we study using macroscopic, and single-channel, recording. We find that gating of the palytoxin-induced channels is regulated by the pump's natural ligands. Thus, external K(+) congeners tend to close, and external Na(+) tends to open, an extracellular gate, whereas ATP acts from the cytoplasmic solution to open an intracellular gate. These gating influences echo the normal ion occlusion and deocclusion reactions that first entrap two extracellular K(+) ions within the interior of the pump (between the two gates) and then release them to the cytoplasmic side in a step accelerated by ATP. These results offer the promise of being able to examine ion occlusion and deocclusion steps at the microscopic level in single Na(+)/K(+) pump molecules.
Artigas
Pablo
P
Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, New York 10021, USA.
Gadsby
David C
DC
D000818:Animals; D006168:Guinea Pigs; D006352:Heart Ventricles; D015640:Ion Channel Gating / Q000502:physiology*; D007473:Ion Channels / Q000502:physiology*; D008954:Models, Biological; D032342:Muscle Cells / Q000187:drug effects / Q000502:physiology*; D037342:Pertussis Toxin / Q000494:pharmacology; D011487:Protein Conformation; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000378:metabolism*
D007473:Ion Channels; D037342:Pertussis Toxin; D000254:Sodium-Potassium-Exchanging ATPase