Adenine nucleotide transport in sonic submitochondrial particles. Kinetic properties and binding of specific inhibitors

Lauquin, Guy J.‐M.; Villiers, Christian; Michejda, J.; Hryniewiecka, L.; Vignais, Pierre V.

doi:10.1016/0005-2728(77)90219-5

Cited by 62 publications

(32 citation statements)

References 20 publications

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“…2B). The transition from uncompetitive to mixed-type inhibition can be attributed to this effect on membrane potential, because similar results were observed when the effects of uncouplers like carbonylcyanide-p-trifluoromethoxyphenyl hydrazone on adenine nucleotide transport in submitochondrial particles were studied (Lauquin et al, 1977).…”

Section: Downloaded Fromsupporting

confidence: 62%

All-trans-Retinoic Acid Binds to and Inhibits Adenine Nucleotide Translocase and induces Mitochondrial Permeability Transition

et al. 2003

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We investigated the effects of retinoic acids on mitochondrial permeability transition (MPT) measured as changes in rhodamine 123 fluorescence from both isolated heart mitochondria and HeLa cells. We report that all-trans-retinoic acid (atRA), 9-cis-retinoic acid, and 13-cis-retinoic acid induce a drop in mitochondrial membrane potential in isolated mitochondria. The atRA effect was done through the induction of MPT because it was dependent on Ca 2ϩ , in a synergic mechanism, and inhibited by cyclosporin A (CsA). Furthermore, atRA also opened MPT in vivo, because treatment of HeLa cells with atRA results in a CsA-sensitive drop of mitochondrial membrane potential. We demonstrated for the first time that retinoic acids inhibit adenine nucleotide translocase (ANT) activity in heart and liver mitochondria. Kinetic studies revealed atRA as an uncompetitive inhibitor of ANT. Photoaffinity labeling of mitochondrial proteins with [3 H]atRA demonstrated the binding of a 31-kDa protein to atRA. This protein was identified as ANT because the presence of carboxyatractyloside, a specific ANT inhibitor, prevented labeling. The specific photolabeling of ANT was also prevented in a concentration-dependent manner by nonlabeled atRA, whereas palmitic acid was ineffective. This study indicates that specific interaction between atRA and ANT takes place regulating MPT opening and adenylate transport. These observations establish a novel mechanism for atRA action, which could control both energetic and apoptotic mitochondrial processes in situations such as retinoic acid treatment.Adenine nucleotide translocase (ANT) is an integral membrane protein of 31 kDa located in mitochondria that catalyzes nucleotide exchange between mitochondria and cytosol. It simultaneously provides ADP for oxidative phosphorylation and ATP to the cytosol for fueling. The existence of two interconvertible conformations of ANT in the mitochondrial membrane has been postulated. The two translocation states of the carrier that correspond to the two conformations are distributed over the cytosol-facing state and matrix-facing state in the presence of ADP or ATP (for reviews, see Klingenberg, 1989;Brandolin et al., 1993).

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Section: Downloaded Fromsupporting

confidence: 62%

All-trans-Retinoic Acid Binds to and Inhibits Adenine Nucleotide Translocase and induces Mitochondrial Permeability Transition

et al. 2003

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“…In order to assess this possibility submitochondrial particles were treated with either carboxyatractylate (3 nmol/ mg of particle protein) or bongkrekic acid (at a final concentration of 10 pM) [28]. Addition to the particles of either adenine-nucleotide-translocator inhibitor had no effect on the size of Ap and AC,, thus demonstrating that mitochondrial contamination was not responsible for any of the observed ATP synthesis (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Clarification of Factors Influencing the Nature and Magnitude of the Protonmotive Force in Bovine Heart Submitochondrial Particles

1981

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The magnitude of the protonmotive force, and its division between pH gradient and membrane potential components has been further characterised in submitochondrial particles. In a reaction medium containing sucrose for osmotic support and 4-(2-hydroxyethyl)-l -piperazineethanesulfonate (Hepes) as buffer, with succinate as substrate, the total protonmotive force reached a maximum value of 245 mV. The presence of CI-enhanced the pH gradient with a partial but not fully compensating decrease in the membrane potential. When submitochondrial particles were suspended in a medium of low osmolarity consisting of phosphoric acid neutralised with Tris, again with succinate as substrate, the protonmotive force was lower and did not exceed 185 mV, and the pH gradient coniponent was equivalent to 25 mV or less.The final phosphorylation potential, AC,, maintained by the particles was higher in the phosphate/Tris medium (46-47.7 kJ mol-.') than in the sucrose/ Hepes/KCI medium (43.7 kJ rnol -'). Thus, comparison of the phosphorylation potential with the protonmotivc force would suggest that the mechanistic stoichiometry H'/ATP (H+ translocated per molecule of ATP synthesied) for the ATPase enzyme is 3 in the former medium and 2 in the latter, which might be taken to indicate two different types of mechanism required for ATP synthesis. However it is questioned whether a comparison of the protonmotive force with AC, in terms of equilibrium thermodynamics ought not to be complemented by analysis in terms of linear non-equilibrium thermodynamics. The latter treatment shows that it is possible to estimate only a value for the product of a phenomenological stoichiometry and the degree of coupling, which can be variable, but not the mechanistic stoichiometry. This treatment can also rationalise the observation of the higher AG, in reaction conditions where the lower values for Ap are estimated. Irrespective of possible explanations, the data show how an unprejudiced choice of reaction conditions can lead to different conclusions about the relationship between the phosphorylation potential and the protonmotive force.Although the acceptance of a chemiosmotic mechanism for membrane-energy transductions is widespread, the problem of quantitatively determining the thermodynamic capacity of the intermediate in chemiosmosis, the protonmotive force, remains.In previous experiments with submitochondrial particles we encountered the somewhat unexpected result that a higher protonmotive force was detected in a reaction mixture that contained sucrose, Hepes and KC1, compared with a reaction mixture that contained phosphoric acid neutralised with Tris base as the main ingredient [1,2]. The lower value for the protonmotive force found in the Pi/Tris medium was not paralleled by a lower value for the phosphorylation potential, so that on a thermodynamic basis a greater number of protons was apparently required per molecule of ATP synthesised in the P,/Tris medium [I], and also the lower protonmotive force could not be attributed to an uncouplingeffe...

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“…The carrier has a turnover number of about 10 s -1 at 18 o C and activation energy of about 45 kJ.mol -1 [5]. The K m values for ADP and ATP are very much dependent on the conditions used for their measurement, and values between 1-100 µM have been reported [5,42,43,[43][44][45][46][47][48]. The nature of the translocation cycle is also debated.…”

Section: Kinetic Models For Ant Mediated Transportmentioning

confidence: 99%

The Adenine Nucleotide Translocase: A Central Component of the Mitochondrial Permeability Transition Pore and Key Player in Cell Death

Halestrap

Brenner²

2003

CMC

446

329

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In addition to its normal function, the adenine nucleotide translocase (ANT) forms the inner membrane channel of the mitochondrial permeability transition pore (MPTP). Binding of cyclophilin-D (CyP-D) to its matrix surface (probably on Pro 61 on loop 1) facilitates a calcium-triggered conformational change converting it from a specific transporter to a non-specific pore. The voltage dependent anion channel (VDAC) binds to the outer face of the ANT, at contact sites between the inner and outer membranes, and together VDAC, ANT and CyP-D probably represent the minimum MPTP configuration. The evidence for this is critically reviewed as is the structure and molecular mechanism of the carrier in its normal physiological mode. This provides helpful insights into MPTP regulation by adenine nucleotides, membrane potential and ANT ligands such as carboxyatractyloside and bongkrekic acid. Oxidative stress activates the MPTP by glutathione-mediated cross-linking of Cys 159 and Cys 256 on matrixfacing loops of the ANT that inhibits ADP binding and enhances CyP-D binding. Molecular modeling of the loop containing the ADP binding site suggests an arrangement of aspartate and glutamate residues that may provide a calcium binding site. There are other proteins that may bind to the ANT, modulating MPTP opening and hence cell death. These included members of the Bax/Bcl-2 family (both oncoproteins and tumor suppressors) and viral proteins. Vpr from HIV-1 can bind to ANT and convert it into a pro-apoptotic pore, whereas vMIA from cytomegalovirus interacts to inhibit opening. Thus the ANT may provide a molecular link between physiopathological mechanisms of infection and the regulation of MPTP function and so represents a potential therapeutic target.

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Adenine nucleotide transport in sonic submitochondrial particles. Kinetic properties and binding of specific inhibitors

Cited by 62 publications

References 20 publications

All-trans-Retinoic Acid Binds to and Inhibits Adenine Nucleotide Translocase and induces Mitochondrial Permeability Transition

All-trans-Retinoic Acid Binds to and Inhibits Adenine Nucleotide Translocase and induces Mitochondrial Permeability Transition

Clarification of Factors Influencing the Nature and Magnitude of the Protonmotive Force in Bovine Heart Submitochondrial Particles

The Adenine Nucleotide Translocase: A Central Component of the Mitochondrial Permeability Transition Pore and Key Player in Cell Death

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