Structural perturbation of the a3-CuB site in mitochondrial cytochrome c oxidase by alcohol solvents.
Ethanol has been observed to cause a perturbation of the catalytic center of the major respiratory protein cytochrome c oxidase. These effects were examined by Fourier transform infrared spectroscopy of carbon monoxide complexes of cytochrome a3Fe and of CuB formed by low-temperature photodissociation of the a3FeCO complex. Carbon monoxide binds to reduced cytochrome oxidase in two major structural forms, alpha and beta, both of which are altered by ethanol. In the absence of ethanol, 15-22% of the total cytochrome oxidase in beef heart mitochondria was observed as beta-forms. Ethanol addition caused a concentration-dependent elimination of the beta-forms with 40% disappearing at 0.05 M (0.23%) ethanol, a concentration that can readily be achieved in the blood of intoxicated individuals. At 0.5 M (2.3%) ethanol and above, almost no beta-forms were detectable. The alpha-CuBCO absorption normally splits into two bands at temperatures below 40 K. This effect was decreased in the presence of ethanol and eliminated by high ethanol concentrations. It appears that ethanol increases the structural fluctuations at the active site of the enzyme, analogous to the effects of increased temperature. There was an 8-10% decrease in the maximum rate of oxygen reduction by mitochondrial cytochrome oxidase in 0.05 M ethanol at 24 degrees C, while higher concentrations of ethanol caused no further inhibition. This is the first demonstration that alpha- and beta-forms of cytochrome c oxidase can be modified by an externally added reagent. Changes in the spectra of alpha-CuBCO in the presence of 50% (v/v) ethylene glycol were quite striking, but variable.(ABSTRACT TRUNCATED AT 250 WORDS)
Fiamingo
F G
FG
Department of Physiological Chemistry, Ohio State University, Columbus 43210.
Jung
D W
DW
Alben
J O
JO
D000818:Animals; D001665:Binding Sites; D002248:Carbon Monoxide; D002417:Cattle; D003300:Copper; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology; D066298:In Vitro Techniques; D007501:Iron; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology; D015394:Molecular Structure; D012997:Solvents / Q000494:pharmacology; D013055:Spectrophotometry, Infrared
D012997:Solvents; D000431:Ethanol; D003300:Copper; D002248:Carbon Monoxide; D007501:Iron; D003576:Electron Transport Complex IV