ATP synthesis driven by alpha-keto acid-stimulated alternative oxidase in pea [Pisum sativum] leaf mitochondria
1997
Vianello, A. (University of Udine (Italy)) | Braidot, E. | Petrussa, E. | Macri, F.
The electrical potential difference (Delta-Psi) generation and ATP synthesis due to alpha-keto acid-stimulated alternative oxidase activity in pea leaf mitochondria, energized by malate plus glutamate, was studied. In the absence of alpha-keto acids, Delta-Psi was ca. 60% collapsed by KCN or antimycin A (AA), while the remaining part (ca. 30%) was dissipated by salicylhydroxamic acid (SHAM). The presence of alpha-keto acids (pyruvate or iodoacetate), known to stimulate the alter native oxidase, caused a decrease in the KCN- or AA-sensitive component of Delta-Psi (ca. 30%), which was accompanied by an increase in the SHAM-sensitive one. When mitochondria were energized by exogenous NADH, succinate or duroquinol, Delta-Psi was completely collapsed by KCN or AA, either in the presence or absence of alpha-keto acids. ADP partially collapsed the SHAM-sensitive component of Delta-Psi, and oligomycin restored this dissipation. In the presence of pyruvate or iodoacetate, ATP content in KCN-treated mitochondria was ca. 40% of control, while that in SHAM plus KCN-inhibited mitochondria was negligible. ADP stimulated oxygen uptake only in the presence of KCN (respiratory control ratio = 1.7), but not in the presence of KCN plus SHAM. It is concluded that alpha-keto acids, stimulating the oxidation of NAD-linked substrates through the alternative oxidase, lead to an increase in the SHAM-sensitive component of Delta-Psi via complex I, thus increasing the contribution of this pathway to ATP synthesis when the cytochrome oxidase is restricted
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