期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 97, 期 -, 页码 501-512出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2016.06.014
关键词
Mitochondria; ROS; Krebs cycle; Pyruvate dehydrogenase; 2-Oxoglutarate dehydrogenase; Cardiac; Liver
资金
- Research and Development Corporation of Newfoundland [5404.1976.101]
- Memorial University MUCEP career development fund
Pyruvate dehydrogenase (Pdh) and 2-oxoglutarate dehydrogenase (Ogdh) are vital for Krebs cycle metabolism and sources of reactive oxygen species (ROS). O-2(center dot-)/H2O2 formation by Pdh and Ogdh from porcine heart were compared when operating under forward or reverse electron transfer conditions. Comparisons were also conducted with liver and cardiac mitochondria. During reverse electron transfer (RET) from NADH, purified Ogdh generated similar to 3-3.5 x more O-2(center dot-)/H2O2 in comparison to Pdh when metabolizing 0.5-10 mu M NADH. Under forward electron transfer (FET) conditions Ogdh generated similar to 2-4 x more O-2(center dot-)/H(2)O(2)than Pdh. In both liver and cardiac mitochondria, Ogdh displayed significantly higher rates of ROS formation when compared to Pdh. Ogdh was also a significant source of ROS in liver mitochondria metabolizing 50 mu M and 500 mu M pyruvate or succinate. Finally, we also observed that DTT directly stimulated O-2(center dot-)/H2O2 formation by purified Pdh and Ogdh and in cardiac or liver mitochondria in the absence of substrates and cofactors. Taken together, Ogdh is a more potent source of ROS than Pdh in liver and cardiac tissue. Ogdh is also an important ROS generator regardless of whether pyruvate or succinate serve as the sole source of carbon. Our observations provide insight into the ROS generating capacity of either complex in cardiac and liver tissue. The evidence presented herein also indicates DTT, a reductant that is routinely added to biological samples, should be avoided when assessing mitochondrial O-2(center dot-)/H2O2 production. (C) 2016 Elsevier B.V. All rights reserved.
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