The modes of action of long chain alkyl compounds on the respiratory chain-linked energy transducing system in submitochondrial particles

Batayneh, N.; Kopacz, Stephan J.; Lee, C.P.

doi:10.1016/0003-9861(86)90752-6

Cited by 23 publications

(11 citation statements)

References 48 publications

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“…An obvious way to overcome an uncertainty in the specific Complex I activity in mitochondria might be to use a detergent to abolish the permeability barrier for NADH. However, the ubiquinone reductase activity of Complex I has been shown to be extremely sensitive to a number of lipophilic compounds including detergents (35)(36)(37), as can be illustrated by strong inhibition of the enzyme by Triton X-100 (36). The permeabilization of intact mitochondria by alamethicin provides a simple procedure for reliable quantitative assay of Complex I without any interference with the substrate translocases and dehydrogenases.…”

Section: The Effect Of Matrix Proteins On Complex I a 7 D Transition mentioning

confidence: 99%

Catalytic Activity of NADH-ubiquinone Oxidoreductase (Complex I) in Intact Mitochondria

Grivennikova

Kapustin

Vinogradov

2001

Journal of Biological Chemistry

101

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The mammalian purified dispersed NADH-ubiquinone oxidoreductase (Complex I) and the enzyme in insideout submitochondrial particles are known to be the slowly equilibrating mixture of the active and de-activated forms (Vinogradov, A. D. (1998) Biochim. Biophys. Acta 1364, 169 -185). We report here the phenomenon of slow active/de-active transition in intact mitochondria where the enzyme is located within its natural environment being exposed to numerous mitochondrial matrix proteins. A simple procedure for permeabilization of intact mitochondria by channel-forming antibiotic alamethicin was worked out for the "in situ" assay of Complex I activity. Alamethicin-treated mitochondria catalyzed the rotenone-sensitive NADH-quinone reductase reaction with exogenousely added NADH and quinoneacceptor at the rates expected if the enzyme active sites would be freely accessible for the substrates. The matrix proteins were retained in alamethicin-treated mitochondria as judged by their high rotenone-sensitive malate-cytochrome c reductase activity in the presence of added NAD ؉ . The sensitivity of Complex I to N-ethylmaleimide and to the presence of Mg 2؉ was used as the diagnostic tools to detect the presence of the de-activated enzyme. The NADH-quinone reductase activity of alamethicin-treated mitochondria was sensitive to neither N-ethylmaleimide nor Mg 2؉ . After exposure to elevated temperature (37°C, the conditions known to induce de-activation of Complex I) the enzyme activity became sensitive to the sulfhydryl reagent and/or Mg 2؉ . The sensitivity to both inhibitors disappeared after brief exposure of the thermally de-activated mitochondria with malate/glutamate, NAD ؉ , and cytochrome c (the conditions known for the turnover-induced reactivation of the enzyme). We conclude that the slow active/ de-active Complex I transition is a characteristic feature of the enzyme in intact mitochondria and discuss its possible physiological significance.In mammalian mitochondria NADH-ubiquinone oxidoreductase (Complex I, coupling Site 1, EC 1.6.99.3) functions as the main entry to the respiratory chain. The enzyme has an extremely complex structure being composed of more than 40 different subunits (1, 2). It contains multiple distinct redox components (FMN, a number of iron-sulfur clusters and tightly bound ubiquinones) operating in unknown sequence of the reactions coupled with vectorial translocation of protons from matrix to intermembraneous space. The functions of a vast majority of the enzyme subunits are not known. Most of the recent studies on Complex I and its simpler procaryotic counterparts (Type 1 NADH dehydrogenases) have focused on their structure (1-4), iron-sulfur clusters location (5, 6), possible mechanism of proton translocation (7-9), and the comparative molecular biology of the enzyme (10 -12).Very little is known about regulatory properties of Complex I. The bovine heart enzyme shows very complex kinetic behavior when assayed in either forward or reverse reactions. Following pioneering observations of Estabr...

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Section: The Effect Of Matrix Proteins On Complex I a 7 D Transition mentioning

confidence: 99%

Catalytic Activity of NADH-ubiquinone Oxidoreductase (Complex I) in Intact Mitochondria

Grivennikova

Kapustin

Vinogradov

2001

Journal of Biological Chemistry

101

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“…Numerous compounds of great diversity in their chemical structure are known to be more or less specific inhibitors of ubiquinone reduction by Complex I [17–19]. Most of them are either very hydrophobic, such as piericidin [20,21] and rotenone [7], or amphiphilic, such as Triton X‐100 [9], n ‐alkyl polyoxyethylenes [22] and fatty acids [23]. Affinity labeling [24,25] and mutational analysis [26–28] suggest that the inhibitors bind to relatively large cavity formed by ND‐1 (NUO‐H), PSST (NUO‐B) and 49 IP (NUO‐D) subunits and located at or close to the interface between the membrane and the peripheral part of Complex I.…”

Section: Discussionmentioning

confidence: 99%

Unidirectional effect of lauryl sulfate on the reversible NADH:ubiquinone oxidoreductase (Complex I)

et al. 2003

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Lauryl sulfate inhibits the v vW W H + + -dependent reverse electron transfer reactions catalyzed by NADH:ubiquinone oxidoreductase (Complex I) in coupled bovine heart submitochondrial particles and in vesicles derived from Paracoccus denitri¢cans. The inhibitor a¡ects neither NADH oxidase (coupled or uncoupled) nor NADH:ferricyanide reductase and succinate oxidase activities at the concentrations that selectively prevent the succinate-supported, rotenone-sensitive NAD + + or ferricyanide reduction. Possible uncoupling e¡ects of the inhibitor are ruled out: in contrast to oligomycin and gramicidin, which increases and decreases the rate of the reverse electron transfer, respectively, in parallel with their coupling and uncoupling e¡ects, lauryl sulfate does not a¡ect the respiratory control ratio. A mechanistic model for the unidirectional e¡ect of lauryl sulfate on the Complex I catalyzed oxidoreduction is proposed. ß

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“…There are few reports on the effect of toxic agents on mitochondrial respiration obtained with various specific substrates of the respiratory chain [26,27]. In the presence of ascorbate + cyt.…”

Section: Discussionmentioning

confidence: 99%

Substrate-Specific Effect of Oleic Acid and Serum from Patients with Reye's Syndrome on Respiratory Control Ratio of Rat Liver Mitochondria

Ogawa

Yoshida

Ogawa

et al. 1988

J. Clin. Biochem. Nutr.

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SummaryThe effects of oleic acid and the serum from two patients with Reye's syndrome (RS-serum) on the respiratory control ratio (RCR) of rat liver mitochondria were studied; the reaction site of each reagent was determined by using various substrates for the respiratory chain. The results were as follows : 1. Pre-incubation of rat liver mitochondri a with oleic acid induced a decrease in RCR when succinate or ascorbate + cytochrome c were employed as substrates. 2. Under the identical conditions, RS-serum induced a significant decrease in RCR with ascorbate + cytochrome c. 3. When KCN was added to the assay medium, both oxygen consumption rate and RCR were decreased as expected in oxidizing ascorbate + cytochrome c. But the effect of KCN was less obvious when pyruvate + L-malate were employed as substrates. The latter phenomenon might suggest that the rate-limiting step in the electron transport system from NADH to O2 resided on the substrate side and not on the side between coenzyme Q and O2.Among the substrates (succinate, pyruvate + L -malate, ascorbate + N,N,N',N'-tetramethyl-phenylenediamine, ascorbate + cytochrome c) tested, the strongest effect was observed when ascorbate + cytochrome c were employed as substrates. These results suggest that both oleic acid and RS-serum showed similar substrate-specific inhibition of RCR, predominantly at the site of cytochrome c oxidase. Thus, a localized effect of oleic acid or RS-serum on the mitochondrial respiratory chain became apparent.

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The modes of action of long chain alkyl compounds on the respiratory chain-linked energy transducing system in submitochondrial particles

Cited by 23 publications

References 48 publications

Catalytic Activity of NADH-ubiquinone Oxidoreductase (Complex I) in Intact Mitochondria

Catalytic Activity of NADH-ubiquinone Oxidoreductase (Complex I) in Intact Mitochondria

Unidirectional effect of lauryl sulfate on the reversible NADH:ubiquinone oxidoreductase (Complex I)

Substrate-Specific Effect of Oleic Acid and Serum from Patients with Reye's Syndrome on Respiratory Control Ratio of Rat Liver Mitochondria

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