Inhibition of the mitochondrial cyclosporin A‐sensitive permeability transition pore by the arginine reagent phenylglyoxal

Eriksson, Ove; Fontaine, Éric; Petronilli, Valeria; Bernardi, Paolo

doi:10.1016/s0014-5793(97)00549-8

Cited by 29 publications

(17 citation statements)

References 16 publications

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Section: Methodssupporting

confidence: 84%

“…The slight inhibitory effect of PGO on uncoupled respiration has been described previously (22), and BAD was found to have similar effects, although at somewhat higher concentrations (data not shown). Mitochondrial swelling, Ca 2ϩ transport, ⌬⌿, oxygen consumption, and matrix pH were measured as described previously (22,25). Peptidylprolyl cis,trans-isomerase activity was measured spectrophotometrically as described (26), except that the assay medium was supplemented with 1% Nonidet P-40.…”

Section: Methodssupporting

confidence: 84%

“…We conclude that the critical arginines are not located on the translocase. Finally, treatment with PGO and BAD affected energy conservation only marginally since only uncoupled respiration was inhibited (22), explaining why the rate of Ca 2ϩ uptake, but not its extent, is affected by BAD and PGO (Fig. 1).…”

Section: Discussionmentioning

confidence: 97%

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Chemical Modification of Arginines by 2,3-Butanedione and Phenylglyoxal Causes Closure of the Mitochondrial Permeability Transition Pore

Eriksson¹,

Fontaine²,

Bernardi³

1998

Journal of Biological Chemistry

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We have investigated the role of arginine residues in the regulation of the mitochondrial permeability transition pore, a cyclosporin A-sensitive inner membrane channel. Isolated rat liver mitochondria were treated with the arginine-specific chemical reagent 2,3-butanedione or phenylglyoxal, followed by removal of excess free reagent. After this treatment, mitochondria accumulated Ca 2؉ normally, but did not undergo permeability transition following depolarization, a condition that normally triggers opening of the permeability transition pore. Inhibition by 2,3-butanedione and phenylglyoxal correlated with matrix pH, suggesting that the relevant arginine(s) are exposed to the matrix aqueous phase. Inhibition by 2,3-butanedione was potentiated by borate and was reversed upon its removal, whereas inhibition by phenylglyoxal was irreversible. Treatment with 2,3-butanedione or phenylglyoxal after induction of the permeability transition by Ca 2؉ overload resulted in pore closure despite the presence of 0.5 mM Ca 2؉. At concentrations that were fully effective at inhibiting the permeability transition, these arginine reagents (i) had no effect on the isomerase activity of cyclophilin D and (ii) did not affect the rate of ATP translocation and hydrolysis, as measured by the production of a membrane potential upon ATP addition in the presence of rotenone. We conclude that reaction with 2,3-butanedione and phenylglyoxal results in a stable chemical modification of critical arginine residue(s) located on the matrix side of the inner membrane, which, in turn, strongly favors a closed state of the pore.Mammalian mitochondria contain an inner membrane channel, the permeability transition pore (PTP) 1 , that, when fully open, permits free diffusion of solutes with a molecular mass of up to ϳ1500 Da (1, 2). The PTP is controlled by several ligands as well as by the ⌬⌿ across the inner membrane; high values of ⌬⌿ favor the closed state, whereas a decrease in ⌬⌿ may increase the open probability (3). High matrix Ca 2ϩ concentrations, P i , and oxidation of intramitochondrial pyridine nucleotides promote pore opening, whereas ADP, H ϩ , and CsA cause inhibition (1, 2). CsA binds to mitochondrial matrix CyP-D (4), which may cause pore inhibition by releasing CyP-D from its putative binding sites on the pore (5, 6). Based on the effect of atractyloside and bongkrekate on the permeability transition (7) and based on the finding that the ANT may form a Ca 2ϩ -dependent large conductance channel in phospholipid bilayers (8), it has been suggested that the pore may be formed by the ANT (7).The physiological function of the permeability transition is not known, but circumstantial evidence suggests that it is involved in Ca 2ϩ homeostasis (9, 10) and linked to cell death. This notion has gained momentum with the findings that (i) Bax and Bcl-XL , two channel-forming proteins of the Bcl-2 superfamily localized mainly to mitochondria (11,12), may affect the PTP and therefore release of the apoptosis-inducing factor, a mitochondrial caspa...

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Section: Methodssupporting

confidence: 84%

Section: Methodssupporting

confidence: 84%

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Chemical Modification of Arginines by 2,3-Butanedione and Phenylglyoxal Causes Closure of the Mitochondrial Permeability Transition Pore

Eriksson¹,

Fontaine²,

Bernardi³

1998

Journal of Biological Chemistry

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“…Isolation of Mitochondria-Liver mitochondria from male Wistar rats were prepared as described (18). The freshly isolated mitochondria (1 mg of protein/ml) were preincubated in modification medium with or without the individual phenylglyoxal derivatives for 15 min at ϩ25°C.…”

Section: Methodsmentioning

confidence: 99%

Modification of Permeability Transition Pore Arginine(s) by Phenylglyoxal Derivatives in Isolated Mitochondria and Mammalian Cells

Johans

Milanesi

Franck

et al. 2005

Journal of Biological Chemistry

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Methylglyoxal and synthetic glyoxal derivatives react covalently with arginine residue(s) on the mitochondrial permeability transition pore (PTP). In this study, we have investigated how the binding of a panel of synthetic phenylglyoxal derivatives influences the opening and closing of the PTP. Using both isolated mitochondria and mammalian cells, we demonstrate that the resulting arginine-phenylglyoxal adduct can lead to either suppression or induction of permeability transition, depending on the net charge and hydrogen bonding capacity of the adduct. We report that phenylglyoxal derivatives that possess a net negative charge and/or are capable of forming hydrogen bonds induced permeability transition. Derivatives that were overall electroneutral and cannot form hydrogen bonds suppressed permeability transition. When mammalian cells were incubated with low concentrations of negatively charged phenylglyoxal derivatives, the addition of oligomycin caused a depolarization of the mitochondrial membrane potential. This depolarization was completely blocked by cyclosporin A, a PTP opening inhibitor, indicating that the depolarization was due to PTP opening. Collectively, these findings highlight that the target arginine(s) is functionally linked with the opening/closing mechanism of the PTP and that the electric charge and hydrogen bonding of the resulting arginine adduct influences the conformation of the PTP. These results are consistent with a model where the target arginine plays a role as a voltage sensor.

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“…6B. CysA inhibits MPT pore opening by interaction with the pore constituent cyclophilin D and has been widely used as a sensitive molecular probe for the involvement of MPT pore opening in cell death pathways (Eriksson et al, 1997). Indeed, CysA pretreatment exerted significant cell protection against induction of apoptosis by DMC.…”

Section: Resultsmentioning

confidence: 99%

Antimelanoma Activity of Apoptogenic Carbonyl Scavengers

Wondrak

Jacobson

2005

J Pharmacol Exp Ther

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Therapeutic induction of apoptosis is an important goal of anticancer drug design. Cellular carbonyl stress mediated by endogenous reactive carbonyl species (RCS) such as glyoxal and methylglyoxal (MG) affects proliferative signaling and metastasis of human tumor cells. Recent research suggests that RCS produced constitutively during increased tumor cell glycolysis may be antiapoptotic survival factors and thus represent a novel molecular target for anticancer intervention. Here, we demonstrate the tumor cell-specific apoptogenicity of carbonyl scavengers, which act by covalently trapping RCS, against human (A375, G361, and LOX) and murine (B16) melanoma cell lines. A structure-activity relationship study identified nucleophilic carbonyl scavenger pharmacophores as the functional determinants of apoptogenic antimelanoma activity of structurally diverse agents such as 3,3-dimethyl-D-cysteine and aminoguanidine. Previous work has demonstrated that covalent adduction of protein-arginine residues in the mitochondrial permeability transition (MPT) pore and heat shock protein 27 by intracellular MG produced in tumor cell glycolysis inhibits mitochondrial apoptosis and enhances cancer cell survival. Indeed, in various melanoma cell lines, carbonyl scavenger-induced apoptosis was antagonized by pretreatment with the membrane-permeable RCS phenylglyoxal (PG). Carbonyl scavenger-induced apoptosis was associated with early loss of mitochondrial transmembrane potential, and cyclosporin A antagonized the effects of carbonyl scavengers, suggesting a causative role of MPT pore opening in carbonyl scavenger apoptogenicity. Consistent with RCS inhibition of mitochondrial apoptosis in melanoma cells, staurosporine-induced apoptosis also was suppressed by PG pretreatment. Our results suggest that carbonyl scavengers acting as direct molecular antagonists of RCS are promising apoptogenic prototype agents for antimelanoma drug design.

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Inhibition of the mitochondrial cyclosporin A‐sensitive permeability transition pore by the arginine reagent phenylglyoxal

Cited by 29 publications

References 16 publications

Chemical Modification of Arginines by 2,3-Butanedione and Phenylglyoxal Causes Closure of the Mitochondrial Permeability Transition Pore

Chemical Modification of Arginines by 2,3-Butanedione and Phenylglyoxal Causes Closure of the Mitochondrial Permeability Transition Pore

Modification of Permeability Transition Pore Arginine(s) by Phenylglyoxal Derivatives in Isolated Mitochondria and Mammalian Cells

Antimelanoma Activity of Apoptogenic Carbonyl Scavengers

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