EFFECT OF CALCIUM-IONS AND INHIBITORS ON INTERNAL NAD(P)H DEHYDROGENASES IN PLANT-MITOCHONDRIA

被引:46
作者
RASMUSSON, AG [1 ]
MOLLER, IM [1 ]
机构
[1] UNIV LUND, DEPT PLANT PHYSIOL, BOX 7007, S-22007 LUND 7, SWEDEN
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1991年 / 202卷 / 02期
关键词
D O I
10.1111/j.1432-1033.1991.tb16415.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Both the external oxidation of NADH and NADPH in intact potato (Solanum tuberosum L. cv. Bintje) tuber mitochondria and the rotenone-insensitive internal oxidation of NADPH by inside-out submitochondrial particles were dependent on Ca2+. The stimulation was not due to increased permeability of the inner mitochondrial membrane. Neither the membrane potential nor the latencies of NAD+-dependent and NADP+-dependent malate dehydrogenases were affected by the addition of Ca2+. The pH dependence and kinetics of Ca2+-dependent NADPH oxidation by inside-out submitochondrial particles were studied using three different electron acceptors: O2, duroquinone and ferricyanide. Ca2+ increased the activity with all acceptors with a maximum at neutral pH and an additional minor peak at pH 5.8 with O2 and duroquinone. Without Ca2+, the activity was maximal around pH 6. The K(m) for NADPH was decreased fourfold with ferricyanide and duroquinone, and twofold with O2 as acceptor, upon addition of Ca2+. The V(max) was not changed with ferricyanide as acceptor, but increased twofold with both duroquinone and O2. Half-maximal stimulation of the NADPH oxidation was found at 3-mu-M free Ca2+ with both O2 and duroquinone as acceptors. This is the first report of a membrane-bound enzyme inside the inner mitochondrial membrane which is directly dependent on micromolar concentrations of Ca2+. Mersalyl and dicumarol, two potent inhibitors of the external NADH dehydrogenase in plant mitochondria, were found to inhibit internal rotenone-insensitive NAD(P)H oxidation, at the same concentrations and in manners very similar to their effects on the external NAD(P)H oxidation.
引用
收藏
页码:617 / 623
页数:7
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