Activation of mitochondrial 2-oxoacid dehydrogenases by thioredoxin.
The regulation of mitochondrial dehydrogenases of 2-oxoacids by thioredoxin is established. It is found that at low NAD+ and saturating concentrations of 2-oxoacids and CoA, inactivation of 2-oxoacid dehydrogenase complexes takes place, preventing NAD+ reduction under such conditions. However, addition of oxidized E. coli thioredoxin to the reaction medium without dithiothreitol allows effective NAD+ reduction at this substrate ratio. Product accumulation curves show that thioredoxin activates the complexes by protecting them from the inactivation observed in the conditions when the complex-bound dihydrolipoate is accumulated. Disappearance of the activatory effect of thioredoxin after its treatment with SH-specific reagents indicates the involvement of the redox-active cysteine couple of thioredoxin in its activation of 2-oxoacid dehydrogenase complexes. The redox-inactive thioredoxin not only shows no activation, but in fact exerts an inhibitory effect. The inhibition manifests the complex formation between SH-modified thioredoxin and dehydrogenase systems, involving amino acid residues of thioredoxin other than cysteine. High efficiency of thioredoxin from E. coli as compared to chloroplast thioredoxin f and glutathione disulfide is revealed. This indicates the importance of specific protein structure also for the influence of the redox-active thioredoxin upon the 2-oxoacid dehydrogenase complexes. The results obtained suggest that these key enzyme systems of mitochondrial metabolism represent previously unidentified targets for the action of mitochondrial thioredoxin, which is known to resemble the E. coli counterpart studies in this work.
Bunik
V
V
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia.
Follmann
H
H
Bisswanger
H
H
D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D000818:Animals; D003545:Cysteine / Q000032:analysis; D004789:Enzyme Activation / Q000187:drug effects; D007655:Ketoglutarate Dehydrogenase Complex / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000201:enzymology; D009097:Multienzyme Complexes / Q000378:metabolism*; D009243:NAD; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D013439:Sulfhydryl Reagents; D013552:Swine; D013879:Thioredoxins / Q000494:pharmacology*
D009097:Multienzyme Complexes; D011768:Pyruvate Dehydrogenase Complex; D013439:Sulfhydryl Reagents; D009243:NAD; D013879:Thioredoxins; D007658:Ketone Oxidoreductases; D007655:Ketoglutarate Dehydrogenase Complex; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D003545:Cysteine