Detection of ros-sensitive thiol proteins by redox-difference gel electrophoresis (redox-dige): Implications for mitochondrial redox signalling. Export

@article{citeulike:1342033, abstract = {Reactive oxygen species (ROS) produced by the mitochondrial respiratory chain can be a redox signal, but whether they affect mitochondrial function is unclear. Here we show that low levels of ROS from the respiratory chain under physiological conditions reversibly modify the thiol redox state of mitochondrial proteins involved in fatty acid and carbohydrate metabolism. As these thiol modifications were specific and occurred without bulk thiol changes, we first had to develop a sensitive technique to identify the small number of proteins modified by endogenous ROS. In this technique, Redox-DIGE (Redox-Difference Gel Electrophoresis), control and redox challenged samples are labelled with different thiol-reactive fluorescent tags and then separated on the same 2D gel, enabling the sensitive detection of thiol redox modifications by changes in the relative fluorescence of the two tags within a single protein spot, followed by protein identification by mass spectrometry. Thiol redox modification affected enzyme activity, suggesting that the reversible modification of enzyme activity by ROS from the respiratory chain may be an important and unexplored mode of mitochondrial redox signalling.}, address = {MRC-Dunn Human Nutrition Unit, Cambridge CB2 2XY.}, author = {Hurd, Thomas R R. and Prime, Tracy A A. and Harbour, Michael E E. and Lilley, Kathryn S S. and Murphy, Michael P P.}, citeulike-article-id = {1342033}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M703591200}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17525152}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17525152}, day = {24}, doi = {10.1074/jbc.M703591200}, issn = {0021-9258}, journal = {J Biol Chem}, keywords = {2d\_gel\_electrophoresis, mitochondria, redox-dige, redox\_signalling, thiol}, month = {May}, posted-at = {2007-05-30 05:53:40}, priority = {2}, title = {Detection of ros-sensitive thiol proteins by redox-difference gel electrophoresis (redox-dige): Implications for mitochondrial redox signalling.}, url = {http://dx.doi.org/10.1074/jbc.M703591200}, year = {2007} }

TY - JOUR ID - citeulike:1342033 L3 - citeulike-article-id:1342033 N2 - Reactive oxygen species (ROS) produced by the mitochondrial respiratory chain can be a redox signal, but whether they affect mitochondrial function is unclear. Here we show that low levels of ROS from the respiratory chain under physiological conditions reversibly modify the thiol redox state of mitochondrial proteins involved in fatty acid and carbohydrate metabolism. As these thiol modifications were specific and occurred without bulk thiol changes, we first had to develop a sensitive technique to identify the small number of proteins modified by endogenous ROS. In this technique, Redox-DIGE (Redox-Difference Gel Electrophoresis), control and redox challenged samples are labelled with different thiol-reactive fluorescent tags and then separated on the same 2D gel, enabling the sensitive detection of thiol redox modifications by changes in the relative fluorescence of the two tags within a single protein spot, followed by protein identification by mass spectrometry. Thiol redox modification affected enzyme activity, suggesting that the reversible modification of enzyme activity by ROS from the respiratory chain may be an important and unexplored mode of mitochondrial redox signalling. JF - J Biol Chem SN - 0021-9258 AD - MRC-Dunn Human Nutrition Unit, Cambridge CB2 2XY. TI - Detection of ros-sensitive thiol proteins by redox-difference gel electrophoresis (redox-dige): Implications for mitochondrial redox signalling. KW - 2d_gel_electrophoresis KW - mitochondria KW - redox-dige KW - redox_signalling KW - thiol AU - Hurd, Thomas R AU - Prime, Tracy A AU - Harbour, Michael E AU - Lilley, Kathryn S AU - Murphy, Michael P PY - 2007/05/24/ UR - http://dx.doi.org/10.1074/jbc.M703591200 ER -