Tight binding of oxaloacetate to succinate dehydrogenase

Priegnitz, Anna; Brzhevskaya, O. N.; Wojtczak, Lech

doi:10.1016/0006-291x(73)90031-4

Cited by 19 publications

(13 citation statements)

References 11 publications

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“…Also of note is that an older report suggested that OAA may bind with extracted SDH in a long-acting fashion that is not easily reversible (23). This does not appear to hold for actively respiring muscle mitochondria, because our data show that restoration of succinate-supported respiration is rapid upon clearing OAA.…”

Section: Inhibition Of Mitochondrial Complex IIcontrasting

confidence: 58%

Oxaloacetic acid mediates ADP-dependent inhibition of mitochondrial complex II–driven respiration

Fink¹,

Bai²,

et al. 2018

Journal of Biological Chemistry

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Edited by John M. DenuWe recently reported a previously unrecognized mitochondrial respiratory phenomenon. When [ADP] was held constant ("clamped") at sequentially increasing concentrations in succinate-energized muscle mitochondria in the absence of rotenone (commonly used to block complex I), we observed a biphasic, increasing then decreasing, respiratory response. Here we investigated the mechanism. We confirmed decades-old reports that oxaloacetate (OAA) inhibits succinate dehydrogenase (SDH). We then used an NMR method to assess OAA concentrations (known as difficult to measure by MS) as well as those of malate, fumarate, and citrate in isolated succinate-respiring mitochondria. When these mitochondria were incubated at varying clamped ADP concentrations, respiration increased at low [ADP] as expected given the concurrent reduction in membrane potential. With further increments in [ADP], respiration decreased associated with accumulation of OAA. Moreover, a low pyruvate concentration, that alone was not enough to drive respiration, was sufficient to metabolize OAA to citrate and completely reverse the loss of succinate-supported respiration at high [ADP]. Further, chemical or genetic inhibition of pyruvate uptake prevented OAA clearance and preserved respiration. In addition, we measured the effects of incremental [ADP] on NADH, superoxide, and H 2 O 2 (a marker of reverse electron transport from complex II to I). In summary, our findings, taken together, support a mechanism (detailed within) wherein succinate-energized respiration as a function of increasing [ADP] is initially increased by [ADP]-dependent effects on membrane potential but subsequently decreased at higher [ADP] by inhibition of succinate dehydrogenase by OAA. The physiologic relevance is discussed.

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Section: Inhibition Of Mitochondrial Complex IIcontrasting

confidence: 58%

Oxaloacetic acid mediates ADP-dependent inhibition of mitochondrial complex II–driven respiration

Fink¹,

Bai²,

et al. 2018

Journal of Biological Chemistry

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“…Most recently the value of 20 mol of -SH groups per mol of flavin has been reported for the purified preparation of succinate dehydrogenase (organic mercurial was used as the reagent) [25]. On the other hand, the active site of the enzyme apparently contains only one sulfhydryl group as judged by stoichiometry between enzyme and tightly bound oxaloacetate, presumably forming the thiosemiacetal bond at the active site [26,27]. Nevertheless the use of MalNEt may give valuable information on the active-site sulfhydryl.…”

Section: Discussionmentioning

confidence: 99%

Reactivity of the Sulfhydryl Groups of Soluble Succinate Dehydrogenase

Vinogradov¹,

Gavrikova²,

Zuevsky³

1976

Eur J Biochem

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Soluble succinate dehydrogenase prepared by butanol extraction reacts with N-ethylmaleimide according to first-order kinetics with respect to both remaining active enzyme and the inhibitor concentration. Binding of the sulfhydryl groups of the enzyme prevents its alkylation by Nethylmaleimide and inhibition by oxaloacetate. A kinetic analysis of the inactivation by alkylating reagent in the presence of succinate or malonate suggests that N-ethylmaleimide acts as a sitcdirected inhibitor. The apparent first-order rate constant of alkylation increases between pH 5.8 and 7.8 indicating a pK, value for the enzyme sulfhydryl group equal to 7.0 at 22 "C in 50 mM Trissulfate buffer. Certain anions (phosphate, citrate, maleate and acetate) decrease the reactivity of the enzyme towards the alkylating reagent. Succinate/phenazine methosulfate reductase activity measured in the presence of a saturating concentration of succinate shows the same pH-dependence as the alkylation rate by N-ethylmaleimide. The mechanism of the first step of succinate oxidation. including a nucleophilic attack of substrate by the active-site sulfhydryl group, is discussed.

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“…The restraint put upon the Krebs cycle during dormancy will, therefore. gradually be released, concomitantly with a restoration of the activity of SDH by succinate (Priegnitz et al 1973) generated in the glyoxylate cycle (Greuter 1972). This will lead to an acceleration of the breakdown of the fatty acid reserves and the conversion of the corresponding metabolites to sugar and CO2, until near depletion of the storage fats, thus allowing an increase in the carbon flow through the glycolytic pathway at the expense of the spore's carbohydrate reserves.…”

Section: Discussionmentioning

confidence: 99%

Self inhibition of the Agaricus bisporus Spore by CO2 and/or γ‐Glulaminyl‐4‐hydroxybenzene and γ‐Gtutaminyl‐3,4‐benzoquinone: A Biochemical Analysis

Rast

Stüyssi

Zobrist

1979

Physiologia Plantarum

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Chemical and enzymological analyses have been carried out to study the mode of action of CO2, in inhibiting germination in Agaricus bisporus and to test the suggestion (Vogel and Weaver, 1972. Exp. Cell Res. 75: 95, Vogel et at, 1974. Am. J. Pathol. 76: 165) that γ‐glutaminyl‐4‐hydroxybenzene (GHB) and/or γ‐glutaminyl‐3,4‐benzoquinone (GBQ) would inhibit the succinate dehydrogenase (SDH) in the mushroom spore and thus represent the causal agent(s) of its dormancy. Evidence is presented that the impairment of SDH is brought about by Ihe CO, fixation product oxaloacelate which is formed in the pyruvate carboxylase reaction, and a mechanism is proposed for the release of the SDH inhibition when activating the spores. These are almost devoid of GHB, whereas fruit bodies of all developmental stages contain considerable amounts of the phenol. GBQ is absent in the spore, and its occurrence in mushroom fruit body extracts appears to be an artifact of extraction. Apart from these analytical results, theoretical reasons are given that vouch against the involvement of the phenolics in the inhibition of SDH in vivo.

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Tight binding of oxaloacetate to succinate dehydrogenase

Cited by 19 publications

References 11 publications

Oxaloacetic acid mediates ADP-dependent inhibition of mitochondrial complex II–driven respiration

Oxaloacetic acid mediates ADP-dependent inhibition of mitochondrial complex II–driven respiration

Reactivity of the Sulfhydryl Groups of Soluble Succinate Dehydrogenase

Self inhibition of the Agaricus bisporus Spore by CO2 and/or γ‐Glulaminyl‐4‐hydroxybenzene and γ‐Gtutaminyl‐3,4‐benzoquinone: A Biochemical Analysis

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