The chloride channel blocker 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) uncouples mitochondria and increases the proton permeability of the plasma membrane in phagocytic cells.
We present evidence that the potent chloride channel blocker NPPB has protonophoric activity in the mitochondria and across the plasma membrane of phagocytic cells. The resting O2 consumption of murine peritoneal macrophages was stimulated up to 2.5-fold in the presence of NPPB, with a K0.5 of 15 microM. The stimulatory effect of NPPB on O2 consumption, like that of the classical protonophore CCCP, was prevented by the mitochondrial respiratory chain inhibitors antimycin A, rotenone or cyanide. NPPB also mediated rheogenic proton transport across the plasma membrane of human neutrophils and macrophages in the direction dictated by the electrochemical proton gradient. As a consequence of its protonophoric activity, NPPB uncoupled mitochondrial ATP synthesis, resulting in partial depletion of cellular ATP. These observations indicate that, at the concentrations frequently used for blockade of anion channels, NPPB acts as an effective protonophore, potentially disturbing cytosolic pH and mitochondrial ATP synthesis.
Lukacs
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GL
Division of Cell Biology, Hospital for Sick Children, Toronto, Canada.
Nanda
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A
Rotstein
O D
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Grinstein
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S
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects; D006801:Humans; D008264:Macrophages / Q000187:drug effects*; D051379:Mice; D008928:Mitochondria / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects*; D009579:Nitrobenzoates / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D010101:Oxygen Consumption / Q000187:drug effects; D011522:Protons; D012402:Rotenone / Q000494:pharmacology; D014475:Uncoupling Agents
D009579:Nitrobenzoates; D011522:Protons; D014475:Uncoupling Agents; D012402:Rotenone; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000968:Antimycin A; D000255:Adenosine Triphosphate