Modulation of oxidative phosphorylation machinery

Authors: Diksha Dani Isao Shimokawa Toshimitsu Komatsu Yoshikazu Higami Uwe Warnken Elham Schokraie Martina Schnölzer Frank Krause Michiru D Sugawa Norbert A Dencher

Publish Date: 2009/11/06

Volume: 11, Issue: 3, Pages: 321-334

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Abstract

Mitochondria being the major source and target of reactive oxygen species ROS play a crucial role during ageing We analyzed ageing and calorie restriction CRinduced changes in abundance of rat liver mitochondrial proteins to understand key aspects behind the ageretarding mechanism of CR The combination of bluenative BN gel system with fluorescence Difference Gel Electrophoresis DIGE facilitated an efficient analysis of soluble and membrane proteins existing as monomers or multiprotein assemblies Changes in abundance of specific key subunits of respiratory chain complexes I IV and V critical for activity and/or assembly of the complexes were identified CR lowered complex I assembly and complex IV activity which is discussed as a molecular mechanism to minimize ROS production at mitochondria Notably the antioxidant system was found to be least affected The GSHGSSG couple could be depicted as a rapid mean to handle the fluctuations in ROS levels led by reversible metabolic shifts We evaluated the relative significance of ROS generation against quenching We also observed parallel and unidirectional changes as effect of ageing and CR in subunits of ATP synthase cytochrome P450 and glutathione Stransferase This is the first report on such ‘putatively hormetic’ ageinganalogous effects of CR besides the ageretarding onesThis work was supported by EC FP6 contract number LSHMCT2004512020 MiMage to NAD and Deutsche Forschungsgemeinschaft grant SFB 472 to NAD and H Seelert We are grateful to Dr Linda Ottoboni Functional Genome analysis DKFZ Heidelberg Germany and Dr Holger Eubel ARC Centre of Excellence in Plant Energy Biology Australia for fruitful discussions on DIGE Kerstin Kammerer and Sabine Fiedler from Functional Proteome Analysis DKFZ Heidelberg Germany for technical assistance in mass spectrometry

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