Respiratory electron transfer activity in an asolectin-isooctane reverse micellar system.
Bovine heart submitochondrial particles (SMP) were solubilized in an asolectin isooctane reverse micellar system and the functionality of the respiratory chain was tested by spectroscopic and amperometric techniques. Electron transfer rate supported by NADH was very slow as evidenced by the low cytochrome reduction levels attained over long incubation periods. In the presence of KCN, NADH caused 34% and 12.5% reduction of the cytochromes aa3 and c, respectively, and negligible reduction of cytochrome b. Supplementation of the system with menadione rose the NADH-dependent reduction of all the cytochromes to levels that were close to the total content. However, no measurable O2 uptake activity took place in the presence of NADH plus menadione, or with ascorbate (or NADH) plus TMPD reducing systems. Therefore, it is suggested that in the organic medium, electron transfer from NADH to O2 is arrested at the terminal oxidase step. Cytochrome oxidase reduced by ascorbate (or NADH) plus TMPD seems to be trapped in its half reduced state (ie, a2+ a3(3+)). Although it is poorly reactive with O2, it can transfer electrons back to cytochrome c and TMPD. The electron transfer block to O2 was overcome when PMS was used instead of TMPD. This seems to be due to the recognized capacity of PMSH2 to carry out simultaneous reduction of both a CuA and a3 CuB redox centers of cytochrome oxidase. The cytochrome oxidase reaction in the organic solvent was highly sensitive to KCN (Ki 1.9 microM) and showed bell-shaped kinetics towards the PMS concentration and a sigmoidal response to water concentration, reaching its maximal turnover number (18 s-1) at 4 mM PMS and 1.1% (v/v) water.(ABSTRACT TRUNCATED AT 250 WORDS)
Escobar
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Unidad de Investigacion Biomedica, Instituto Mexicano del Seguro Social, Mexico, DF.
Escamilla
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D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism; D002417:Cattle; D003573:Cytochrome b Group / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism; D007700:Kinetics; D008823:Micelles; D008929:Mitochondria, Heart / Q000378:metabolism*; D009243:NAD / Q000378:metabolism / Q000494:pharmacology; D009795:Octanes; D010101:Oxygen Consumption; D010713:Phosphatidylcholines; D010743:Phospholipids; D011190:Potassium Cyanide / Q000494:pharmacology; D013367:Submitochondrial Particles / Q000378:metabolism*; D013772:Tetramethylphenylenediamine / Q000378:metabolism; D014812:Vitamin K / Q000494:pharmacology
D003573:Cytochrome b Group; D003574:Cytochrome c Group; D008823:Micelles; D009795:Octanes; D010713:Phosphatidylcholines; D010743:Phospholipids; D009243:NAD; D014812:Vitamin K; C024990:asolectin; D003576:Electron Transport Complex IV; D011190:Potassium Cyanide; D013772:Tetramethylphenylenediamine; D001205:Ascorbic Acid; C045798:2,2,4-trimethylpentane