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PubMed · 10973817

Brain mitochondrial complex I inactivation by oxidative modification.

J Bautista·R Corpas·R Ramos·O Cremades·J F Gutiérrez·S Alegre
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Abstract

In vitro oxidation of the brain mitochondrial complex I by the hydroxyl radical generating system ascorbate/Fe(III)/O(2) has been carried out. Complex I inactivation, by oxidation, has been studied using a method based on the resolution of proteins by blue native polyacrylamide gel electrophoresis (BN-PAGE), followed by total protein quantification by staining with Coomassie brilliant blue, in-gel activity quantification, and quantification of oxidized proteins by labelling with DIG-hydrazide and immunodetection with an anti-DIG-AP. Quantification was carried out by densitometry procedure. Our results show that oxidation is a continuous process, increasing rapidly at the beginning, reaching a plateau after 8 h of incubation. There is practically no inactivation until a threshold value of damage is reached. Below this, the complex activity is resistant to the aggression of oxygen-reactive substances and free radicals, but once the threshold value is passed, activity is lost rapidly.

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AnimalsAscorbic AcidBrainDigoxigeninElectron TransportElectron Transport Complex IElectrophoresis, Polyacrylamide GelHydroxyl RadicalImmunoblottingIronKineticsMitochondriaNADH, NADPH OxidoreductasesOxidantsRatsStaining and LabelingJournal Article

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J Bautista, R Corpas, R Ramos, O Cremades, J F Gutiérrez, S Alegre. 2000-09-07. Brain mitochondrial complex I inactivation by oxidative modification.. https://doi.org/10.1006/bbrc.2000.3388

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