期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 106, 期 -, 页码 302-314出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2017.02.046
关键词
Mitochondria; Pyruvate dehydrogenase; S-glutathionylation; Reactive oxygen species; Redox signaling
资金
- Ignite Research Grant provided by the Research and Development Corporation (RDC) [5404.1976.101]
- Memorial University of Newfoundland University Student Summer Internship Program (USSIP)
- Memorial University of Newfoundland Summer Undergraduate Research Award (SURA)
Pyruvate dehydrogenase (Pdh) is a vital source of reactive oxygen species (ROS) in several different tissues. Pdh has also been suggested to serve as a mitochondrial redox sensor. Here, we report that O-2(center dot-)/H2O2 emission from pyruvate dehydrogenase (Pdh) is altered by S-glutathionylation. Glutathione disulfide (GSSG) amplified O-2(center dot-)/H2O2 production by purified Pdh during reverse electron transfer (RET) from NADH. Thiol oxidoreductase glutaredoxin-2 (Grx2) reversed these effects confirming that Pdh is a target for S-glutathionylation. S-glutathionylation had the opposite effect during forward electron transfer (FET) from pyruvate to NAD(+) lowering O-2(center dot-)/H2O2 production. Immunoblotting for protein glutathione mixed disulfides (PSSG) following diamide treatment confirmed that purified Pdh can be S-glutathionylated. Similar observations were made with mouse liver mitochondria. S-glutathionylation catalysts diamide and disulfiram significantly reduced pyruvate or 2-oxoglutarate driven O-2(center dot-)/H2O2 production in liver mitochondria, results that were confirmed using various Pdh, 2-oxoglutarate dehydrogenase (Ogdh), and respiratory chain inhibitors. Immunoprecipitation of Pdh and Ogdh confirmed that either protein can be S-glutathionylated by diamide and disulfiram. Collectively, our results demonstrate that the S-glutathionylation of Pdh alters the amount of ROS formed by the enzyme complex. We also confirmed that Ogdh is controlled in a similar manner. Taken together, our results indicate that the redox sensing and ROS forming properties of Pdh and Ogdh are linked to S-glutathionylation.
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