Abstract
We had previously suggested that phosphorylation of proteins by mitochondrial kinases regulate the activity of NADH/CoQ oxidoreductase. Initial data showed that pyruvate dehydrogenase kinase (PDK) and cAMP-dependent protein kinase A (PKA) phosphorylate mitochondrial membrane proteins. Upon phosphorylation with crude PDK, mitochondria appeared to be deficient in NADH/cytochrome c reductase activity associated with increased superoxide production. Conversely, phosphorylation by PKA resulted in increased NADH/cytochrome c reductase activity and decreased superoxide formation. Current data confirms PKA involvement in regulating Complex I activity through phosphorylation of an 18 kDa subunit. Beef heart NADH/cytochrome c reductase activity increases to 150% of control upon incubation with PKA and ATP-γ-S. We have cloned the four human isoforms of PDK and purified beef heart Complex I. Incubation of mitochondria with PDK isoforms and ATP did not alter Complex I activity or superoxide production. Radiolabeling of mitochondria and purified Complex I with PDK failed to reveal phosphorylated proteins.
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Maj, M.C., Raha, S., Myint, T. et al. Regulation of NADH/CoQ Oxidoreductase: Do Phosphorylation Events Affect Activity?. J Protein Chem 23, 25–32 (2004). https://doi.org/10.1023/B:JOPC.0000016255.17077.2c
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DOI: https://doi.org/10.1023/B:JOPC.0000016255.17077.2c