Decoupling of the Catalytic and Transport Activities of Complex I from Rhodothermus marinus by Sodium/Proton Antiporter Inhibitor

• Published in: ACS chemical biology Vol. 6; no. 5; pp. 477 - 483
• Main Authors: Batista, Ana P, Marreiros, Bruno C, Pereira, Manuela M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 20-05-2011
• Subjects: Amiloride - analogs & derivatives; Amiloride - pharmacology; Biological Transport - drug effects; Catalysis; Electron Transport Complex I - antagonists & inhibitors; Electron Transport Complex I - metabolism; Hydrogen - metabolism; Hydrogen-Ion Concentration; Ion Transport - drug effects; Membrane Potentials - drug effects; Quinone Reductases - antagonists & inhibitors; Rhodothermus - metabolism; Sodium - metabolism; Sodium-Hydrogen Exchangers - antagonists & inhibitors; Valinomycin - pharmacology
• ISSN: 1554-8929; 1554-8937
• DOI: 10.1021/cb100380y

The energy transduction by complex I from Rhodothermus marinus was addressed by studying the influence of 5-(N-ethyl-N-isopropyl)-amiloride (EIPA) on the activities of this enzyme. EIPA is an inhibitor of both Na+/H+ antiporter and complex I NADH:quinone oxidoreductase activity. We performed studies of NADH:quinone oxidoreductase and H+ and Na+ translocation activities of complex I from R. marinus at different concentrations of EIPA, using inside-out membrane vesicles. We observed that the oxidoreductase activity and both H+ and Na+ transports are inhibited by EIPA. Most interestingly, the catalytic and the two transport activities showed different inhibition profiles. The transports are inhibited at concentrations of EIPA at which the catalytic activity is not affected. In this way the catalytic and transport activities were decoupled. Moreover, the inhibition of the catalytic activity was not influenced by the presence of Na+, whereas the transport of H+ showed different inhibition behaviors in the presence and absence of Na+. Taken together our observations indicate that complex I from R. marinus performs energy transduction by two different processes: proton pumping and Na+/H+ antiporting. The decoupling of the catalytic and transport activities suggests the involvement of an indirect coupling mechanism, possibly through conformational changes.