Membrane Potential-Dependent Inhibition of the Na+,K+-ATPase bypara-Nitrobenzyltriethylammonium Bromide
| Autor: | Joshua R. Berlin, R. Daniel Peluffo |
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| Rok vydání: | 2012 |
| Předmět: |
Bromides
Stereochemistry Sodium-Potassium-Exchanging ATPase Ouabain Membrane Potentials medicine Animals Myocytes Cardiac Enzyme Inhibitors Binding site Na+/K+-ATPase Pharmacology chemistry.chemical_classification Membrane potential Binding Sites Sodium Articles Rats Quaternary Ammonium Compounds Kinetics Transmembrane domain A-site Enzyme chemistry Potassium Molecular Medicine medicine.drug |
| Zdroj: | Molecular Pharmacology. 82:1-8 |
| ISSN: | 1521-0111 0026-895X |
| Popis: | Membrane potential (V(M))-dependent inhibitors of the Na(+),K(+)-ATPase are a new class of compounds that may have inherent advantages over currently available drugs targeting this enzyme. However, two questions remain unanswered regarding these inhibitors: (1) what is the mechanism of V(M)-dependent Na(+),K(+)-ATPase inhibition, and (2) is their binding affinity high enough to consider them as possible lead compounds? To address these questions, we investigated how a recently synthesized V(M)-dependent Na(+),K(+)-ATPase inhibitor, para-nitrobenzyltriethylamine (pNBTEA), binds to the enzyme by measuring the extracellular pNBTEA concentration and V(M) dependence of ouabain-sensitive transient charge movements in whole-cell patch-clamped rat cardiac ventricular myocytes. By analyzing the kinetics of charge movements and the steady-state distribution of charge, we show that the V(M)-dependent properties of pNBTEA binding differ from those for extracellular Na(+) and K(+) binding, even though inhibitor binding is competitive with extracellular K(+). The data were also fit to specific models for pNBTEA binding to show that pNBTEA binding is a rate-limiting V(M)-dependent reaction that, in light of homology models for the Na(+),K(+)-ATPase, we interpret as a transfer reaction of pNBTEA from a peripheral binding site in the enzyme to a site near the known K(+) coordination sites buried within the transmembrane helices of the enzyme. These models also suggest that binding occurs with an apparent affinity of 7 μM. This apparent binding affinity suggests that high-affinity V(M)-dependent Na(+),K(+)-ATPase inhibitors should be feasible to design and test as specific enzyme inhibitors. |
| Databáze: | OpenAIRE |
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