Dihydrolipoic acid activates oligomycin-sensitive thiol groups and increases ATP synthesis in mitochondria

Zimmer, Guido; Mainka, Luise; Krüger, Erika

doi:10.1016/0003-9861(91)90243-c

Cited by 45 publications

(10 citation statements)

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“…Thus, lipoic acid is thought to have restored the environments surrounding the enzyme by lowering peroxidation of membrane lipids. Further, α‐lipoic acid might have prevented other age‐dependent mitochondrial dysfunction and subsequently increased the availability of ATP 10…”

Section: Discussionmentioning

confidence: 99%

Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Arivazhagan

Panneerselvam

2004

Annals of the New York Academy of Sciences

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A convincing link between oxidative stress and neurodegenerative diseases has been found with the knowledge that it actually damages neuronal cells in culture. We analyzed the effect of DL-alpha-lipoic acid on lipofuscin and Na(+)K(+) ATPase in discrete brain regions of young and aged rats. In aged rats, the level of lipofuscin was increased, and the activity of Na(+)K(+)ATPase was decreased. Intraperitoneal administration of lipoic acid to aged rats led to a duration-dependent reduction and elevation in lipofuscin and enzyme activity, respectively, in the cortex, cerebellum, striatum, hippocampus, and hypothalamus of the brain. These results suggest that lipoic acid, a natural metabolic antioxidant, should be useful as a therapeutic tool in preventing neuronal dysfunction in aged individuals.

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Section: Discussionmentioning

confidence: 99%

Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Arivazhagan

Panneerselvam

2004

Annals of the New York Academy of Sciences

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“…However, the possibility remains that another enzyme takes over the normal function of the dihydrolipoamide dehydrogenase when it is absent, although no such precedent is known. Alternatively, dihydrolipoic acid is known to possess antioxidant properties (27), and a role for this cofactor in the repair of membrane thiol groups in mitochondria has been suggested (31). Therefore, the role of lipoic acid and dihydrolipoamide dehydrogenase in the halophilic Archaea may be in protection and repair, and this function may not be evident until the cells are stressed in some oxidative manner.…”

Section: Discussionmentioning

confidence: 99%

Dihydrolipoamide dehydrogenase from the halophilic archaeon Haloferax volcanii: homologous overexpression of the cloned gene

et al. 1996

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The gene encoding dihydrolipoamide dehydrogenase from the halophilic archaeon, Haloferax volcanii, has been subcloned and overexpressed in the parent organism by using the halophilic archaeal rRNA promoter. The recombinant protein has been purified to homogeneity and characterized with respect to its kinetic, molecular, and salt-dependent properties. A dihydrolipoamide dehydrogenase-minus mutant of H. volcanii has been created by homologous recombination with the subcloned gene after insertion of the mevinolin resistance determinant into the protein-coding region. To explore the physiological function of the dihydrolipoamide dehydrogenase, the growth properties of the mutant halophile have been examined. Dihydrolipoamide dehydrogenase catalyzes the oxidation ofIn members of the domains Bacteria and Eucarya, the enzyme normally performs this function as an integral component of the pyruvate, 2-oxoglutarate, and branchedchain 2-oxoacid dehydrogenase multienzyme complexes (19,20) or of the glycine cleavage system (16). In all of these systems, the lipoyl moiety is covalently bound to the enzyme complexes, where its reduction and reoxidation are part of the catalytic cycle.Whereas the role in these complexes is the only established function of dihydrolipoamide dehydrogenase, the enzyme has now been found in a variety of Bacteria, Eucarya, and Archaea in the apparent absence of the multienzyme systems (10,11,23,24,26). Such observations are indicative of a new function for the dehydrogenase (8), although its precise nature is still to be established. Clearly the identification of a new role requires evidence from both molecular genetics and protein structure, and to this end we have chosen to study in detail the dihydrolipoamide dehydrogenase from the moderately halophilic archaeon, Haloferax volcanii.The discovery of dihydrolipoamide dehydrogenase and its substrate lipoic acid in the halophilic Archaea, and of the apparent absence of the 2-oxoacid dehydrogenase complexes in these organisms, is reviewed in references 9 and 12. The gene encoding the enzyme has been cloned and sequenced from H. volcanii (29), and from sequence alignments it is clearly related to the complexed dihydrolipoamide dehydrogenases from Bacteria and Eucarya. In addition, shuttle vectors with antibiotic resistance markers for H. volcanii are now available (13-15) to enable the necessary molecular biological analysis of this halophilic gene and its enzyme product.In this paper, we report the subcloning and expression of the H. volcanii gene, the purification and characterization of the recombinant dihydrolipoamide dehydrogenase, and the creation and growth analysis of a H. volcanii strain that lacks the functional enzyme. MATERIALS AND METHODSReagents and enzymes. Bovine serum albumin, DL-lipoamide, RNase A, lowmelting-point agarose, novobiocin, and polyethylene glycol (PEG) 600 were purchased from Sigma. PEG 600 was purified by the method of Klebe et al. (17) DL-Dihydrolipoamide was prepared by the reduction of DL-lipoamide with NaBH 4 (22); t...

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“…As is becoming evident from the foregoing discussion, in mitochondria, oligomycin -sensitive thiol reactivity occurs to a remarkable extent [1,4,7,8,[18][19][20]33,43,57,58,61,62,64,66]. Such mitochondrial -SH reactivity can be also deduced to working rat hearts in experimentation with MPG [65] and alpha lipoic acid [67].…”

Section: Discussionmentioning

confidence: 99%

“…2). Our previous findings [61][62][63][64]20,21,65] using 2-mercapto-propionylglycine (MPG) and the more recent results using alpha lipoic acid [3,11,34,39,47,50,66,68], could, at least in part, become explained by reversible blockade (R-S-MPG; R-S-LA), of reactive sulfhydryl groups at strategic membrane sites including the ATPsynthase with excessive reversible reactivity of the Fo b subunit preserving mobility and structure changes during ATP synthesis.…”

Section: Discussionmentioning

confidence: 99%

“…Thereafter, the "new" sulfhydryls were characterized by radioactive labeling. For this goal, we used the weakly hydrophilic (−0.47 = log Pow, personal communication Dr. G. Peter, ASTA Medica, Frankfurt) [ 14 C]-labeled lipoic acid, a physiological substance which was previously found to become most interesting due to its antioxidant and thiol-preserving actions [11,34,39,50,66,68]. The studies were performed with lipoic acid [ 14 C] labeled on 7, 8 position.…”

Section: Introductionmentioning

confidence: 99%

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ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

Dünschede¹,

Zwicker

Ackermann

et al. 2003

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Purified ATPsynthase of bovine heart mitochondria has been analyzed for its mobility and reactivity of oligomycin-sensitive sulfhydryl regions in presence of the substrate ADP and oligomycin. Labeling of thiol groups at the hydrophobic F_0 region of the ATPsynthase was increased in the enzyme initially treated with SDS, N-ethylmaleimide and dithiothreitol (modified enzyme). After dialysis or gel permeation the ATPsynthase was treated with [14C] alpha lipoic acid at a molar ratio of 35-85/1 (lipoic acid/ATPsynthase) corresponding to 4-8.6 nmol/mg protein. Under these conditions, ATPase activity of the native enzyme was significantly decreased. After preincubation with ADP, PAGE of the native, [14C] labeled enzyme revealed an increase of radioactivity at a region of 25 kDa deduced to Cys 197 of subunit b. In the modified enzyme the increase in radioactivity was found at 10 kDa. In this context, the sequence Lys-Cys-Ile around Cys 197 of subunit b suggests excessive reactivity of this thiol, as well as ready reversibility by -SH-S-S- interchange. Therefore, previously observed reaction by thiol reagents and antioxidants from outside the mitochondrion can be interpreted with Cys 197 of F0 b. It accounts for sulfhydryl unmasked by binding of ADP at F1.

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Dihydrolipoic acid activates oligomycin-sensitive thiol groups and increases ATP synthesis in mitochondria

Cited by 45 publications

References 23 publications

Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Dihydrolipoamide dehydrogenase from the halophilic archaeon Haloferax volcanii: homologous overexpression of the cloned gene

ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

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Dihydrolipoic acid activates oligomycin-sensitive thiol groups and increases ATP synthesis in mitochondria

Cited by 45 publications

References 23 publications

Alpha‐Lipoic Acid Increases Na+K+ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Alpha‐Lipoic Acid Increases Na+K+ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Dihydrolipoamide dehydrogenase from the halophilic archaeon Haloferax volcanii: homologous overexpression of the cloned gene

ADP‐ and oligomycin‐sensitive redox behavior of F0 b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

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Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

Alpha‐Lipoic Acid Increases Na⁺K⁺ATPase Activity and Reduces Lipofuscin Accumulation in Discrete Brain Regions of Aged Rats

ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid