Catalytic activity of cytochrome oxidase and cytochrome c in apolar solvents containing phospholipids and low amounts of water.
Cytochrome c and cytochrome oxidase, in bovine heart submitochondrial particles and in their purified forms, were transferred to a ternary system that contained phospholipids (10 mg/ml toluene), the apolar solvent toluene, and water at concentrations of 13-15 microliters (high water) and 3 microliters (low water) per milliliter of toluene. When the enzymes were transferred back to an all water system, they exhibited full catalytic capacity. In the low water ternary system, cytochrome c could be reduced by ascorbate introduced via inverted micelles. Also in this system, cytochrome oxidase was reduced by ascorbate and cytochrome c but its oxidation was highly impaired. Data on the kinetics of reduction by ascorbate of cytochrome c and cytochrome oxidase under these conditions are presented. Cytochrome oxidase reduced in the organic solvent by ascorbate failed to form a complex with CO, but formed a complex with cyanide introduced via inverted micelles. The oxidized and the ascorbate-reduced cytochrome oxidase-cyanide complex exhibited a trough at 415 nm and a peak at 433 nm. The extent and rate of formation of the cyanide complex were higher with the reduced form of cytochrome oxidase. To achieve protein-protein interactions (cytochrome c-cytochrome oxidase) in the ternary system, it was necessary to extract the two proteins together. There was no functional interaction when they were extracted separately and mixed. In the high water ternary system reduced cytochrome oxidase was not detected, and it oxidized ascorbate at a higher rate than in the low water system; however, this rate was several orders of magnitude lower than in aqueous media.
Escamilla
E
E
Instituto de Fisiologia Celular, Universidad Nacional Autónoma de México, DF.
Ayala
G
G
de Gómez-Puyou
M T
MT
Gómez-Puyou
A
A
Millán
L
L
Darszon
A
A
D000818:Animals; D001205:Ascorbic Acid; D002248:Carbon Monoxide / Q000494:pharmacology; D002417:Cattle; D003486:Cyanides / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism*; D003576:Electron Transport Complex IV / Q000378:metabolism*; D066298:In Vitro Techniques; D008929:Mitochondria, Heart / Q000201:enzymology; D010084:Oxidation-Reduction; D010743:Phospholipids; D012997:Solvents; D013057:Spectrum Analysis; D013367:Submitochondrial Particles / Q000201:enzymology; D014050:Toluene; D014867:Water
D003486:Cyanides; D003574:Cytochrome c Group; D010743:Phospholipids; D012997:Solvents; D014867:Water; D014050:Toluene; D002248:Carbon Monoxide; D003576:Electron Transport Complex IV; D001205:Ascorbic Acid