Inhibitory effects of two structurally related carbocyanine laser dyes on the activity of bovine heart mitochondrial and Paracoccus denitrificans NADH-ubiquinone reductase. Evidence for a rotenone-type mechanism.
Two cationic, lipophilic laser dyes, 1,1',3,3,3',3'-hexamethylindodicarbocyanine iodide (HIDC) and 1,1',3,3,3',3'-hexamethylindotricarbocyanine iodide (HITC), inhibit bovine heart mitochondrial and Paracoccus denitrificans NADH oxidase activities. The mitochondrial I50 values were 0.5 microM (HIDC) and 1.2 microM (HITC), and the P. denitrificans I50 values 1.2 microM (HIDC) and 1.5 microM (HITC). Neither succinate nor cytochrome oxidase (EC 1.9.3.1) activities were inhibited significantly by either compound, localizing the site of inhibition to the segment of each electron transport chain between NADH and ubiquinone. With submitochrondrial particles (SMP), NADH-dependent reduction of menadione, duroquinone and coenzyme Q1 was inhibited markedly (HIDC was the more potent inhibitor). Using purified complex I, only NADH-dependent reduction of duroquinone and coenzyme Q1 was inhibited markedly (HIDC was the more potent inhibitor) and reduction of menadione was inhibited slightly. With P. denitrificans membrane vesicles, NADH-dependent reduction of menadione, juglone, and coenzyme Q1 was inhibited slightly and duroquinone reduction was inhibited markedly. Membrane-dependent interactions appear to be involved, since the compounds were more inhibitory with membrane preparations than with complex I. The mechanism of inhibition (except for the HIDC effect on coenzyme Q1 reduction with P. denitrificans) appeared to be through the interaction of dye with the rotenone site on NADH-ubiquinone reductase (EC 1.6.99.3), since rotenone-insensitive preparations of complex I and P. denitrificans membrane vesicles were also insensitive to HIDC and HITC inhibition.
Anderson
W M
WM
Indiana University School of Medicine, Northwest Center for Medical Education, Gary 46408.
Chambers
B B
BB
Wood
J M
JM
Benninger
L
L
D000818:Animals; D002232:Carbocyanines / Q000494:pharmacology*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D004579:Electron Transport / Q000187:drug effects; D005456:Fluorescent Dyes / Q000494:pharmacology*; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000201:enzymology; D016660:NAD(P)H Dehydrogenase (Quinone); D010231:Paracoccus denitrificans / Q000187:drug effects* / Q000201:enzymology; D011808:Quinone Reductases / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology*
D002232:Carbocyanines; D005456:Fluorescent Dyes; D012402:Rotenone; C053313:1,1',3,3,3',3'-hexamethylindotricarbocyanine; C067566:1,1',3,3,3',3'-hexamethylindodicarbocyanine; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases