Allosteric regulation of rat testis mitochondrial aldehyde dehydrogenase by capronaldehyde and magnesium ion.
1. The influence of Mg2+ on the kinetic behaviour of mitochondrial aldehyde dehydrogenase from rat testis has been investigated using capronaldehyde as substrate. 2. The kinetic data, obtained by numerical analysis of the progress curves of aldehyde oxidation, were fitted to a modified version of the Monod-Wyman-Changeux model and the fitting procedure resulted in a good correspondence between theoretical and experimental reaction rates over a wide range of capronaldehyde and Mg2+ concentrations. 3. According to the model, the tetrameric enzyme is in equilibrium between two conformational states R and T which display comparable affinities for capronaldehyde (the dissociation constants are 0.17 and 0.3 microM, respectively), but different catalytic power (VT = 2VR). The T state can bind with lower affinity a second molecule of aldehyde (K = 2.5 microM). 4. Mg2+ stabilizes the T state (the dissociation constants for the R and T states are 2.2 and 0.12 mM, respectively) and acts as a strong activator of the R state, but as a weak inhibitor of the T state. In the absence of substrates and Mg2+, the R<-->T equilibrium favors the R state ([T]/[R] = 0.16). 5. The model is able to predict the kinetic behaviour also when the NAD+ concentrations are not saturating and when inhibitory effects by NADH are taken into account.
Bedino
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S
Dipartimento di Medicina e Oncologia Sperimentale, Università di Torino, Italy.
Testore
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G
D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000494:Allosteric Regulation; D000818:Animals; D007700:Kinetics; D008274:Magnesium / Q000502:physiology*; D008297:Male; D008928:Mitochondria / Q000201:enzymology*; D008954:Models, Biological; D009243:NAD / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012044:Regression Analysis; D013737:Testis / Q000201:enzymology* / Q000648:ultrastructure
D000447:Aldehydes; D009243:NAD; C010463:n-hexanal; D000444:Aldehyde Dehydrogenase; D008274:Magnesium