title: Assessment of uncoupling by amiloride analogs.
abstract: The amiloride analogs N5-methyl-N5-isobutylamiloride, N5-ethyl-N5-isopropylamiloride, and N5,N5-hexamethyleneamiloride are frequently used to investigate NaH exchange on the premise that they are highly specific inhibitors of the NaH-antiporters. We assessed the relative protonophoric activity of these compounds in reconstituted and native membrane vesicles, using acridine orange fluorescence to measure intravesicular pH. All the compounds tested were found to be potent protonophores at concentrations which are normally used to demonstrate inhibition of NaH exchange. Uncoupling was dependent on both the pH of the assay system and the total amount of lipid present. At the pH optima, which lay in a range from 7.5 to 8.5, these amiloride analogs were more potent uncouplers than the classical protonophore carbonyl cyanide m-chlorophenylhydrazone. Therefore, extreme care must be taken in the interpretation of results obtained using these or similar derivatives of amiloride.
authors.affiliation: Department of Clinical Pharmacology, University of Berne, Switzerland.
authors.forename: K
authors.initials: K
authors.lastname: Davies
authors.forename: M
authors.initials: M
authors.lastname: Solioz
mesh_terms: D000165:Acridine Orange; D000584:Amiloride / Q000031:analogs & derivatives*; D000643:Ammonium Chloride; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors*; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000494:pharmacology; D008081:Liposomes / Q000378:metabolism; D011522:Protons; D017923:Sodium-Hydrogen Exchangers; D013050:Spectrometry, Fluorescence; D014475:Uncoupling Agents / Q000494:pharmacology*
chemical_list: D002352:Carrier Proteins; D008055:Lipids; D008081:Liposomes; D011522:Protons; D017923:Sodium-Hydrogen Exchangers; D014475:Uncoupling Agents; D000643:Ammonium Chloride; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000584:Amiloride; D000165:Acridine Orange