Phosphate is essential for inhibition of the mitochondrial permeability transition pore by cyclosporin A and by cyclophilin D ablation - PubMed (original) (raw)

Phosphate is essential for inhibition of the mitochondrial permeability transition pore by cyclosporin A and by cyclophilin D ablation

Emy Basso et al. J Biol Chem. 2008.

Abstract

Energized mouse liver mitochondria displayed the same calcium retention capacity (a sensitive measure of the propensity of the permeability transition pore (PTP) to open) irrespective of whether phosphate, arsenate, or vanadate was the permeating anion. Unexpectedly, however, phosphate was specifically required for PTP desensitization by cyclosporin A (CsA) or by genetic inactivation of cyclophilin D (CyP-D). Indeed, when phosphate was replaced by arsenate, vanadate, or bicarbonate, the inhibitory effects of CsA and of CyP-D ablation on the PTP disappeared. After loading with the same amount of Ca(2+) in the presence of arsenate or vanadate but in the absence of phosphate, the sensitivity of the PTP to a variety of inducers was identical in mitochondria from wild-type mice, CyP-D-null mice, and wild-type mice treated with CsA. These findings call for a reassessment of conclusions on the role of the PTP in cell death that are based on the effects of CsA or of CyP-D ablation.

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Figures

FIGURE 1.

Effect of anions on the mitochondrial permeability transition. A, the incubation medium was supplemented with 1 m

Asi (trace a), 1 m

Vi (trace b), or 1 m

Pi (trace c). The conditions were as follows: 2-ml final volume, pH 7.4, and 25 °C. Where indicated, 1 mg of mouse liver mitochondria (M) was added, followed by a train of Ca2+ pulses of 10 μ

each (arrows).B, the experimental conditions were as described for A, except that the medium contained 0.1 m

Pi plus the indicated concentrations of Asi (▴), Vi (▪), and Pi (•). The CRC is expressed in nanomoles of Ca2+/mg of protein.

FIGURE 2.

Desensitization of the PTP by CsA or by CyP-D ablation depends on Pi. Where indicated (M), 1 mg of wild-type mitochondria treated with 0.8 μ

CsA (traces a–c) or of CyP-D-null mitochondria (_traces a_′_–c_′) was added to medium containing 1 m

Asi (traces a and _a_′), 1 m

Vi (traces b and _b_′), or 1 m

Pi (traces c and c_′) under conditions otherwise identical to those described in the legend to Fig. 1_A.

FIGURE 3.

Desensitization of the PTP by CsA or by CyP-D ablation depends on Pi. A, wild-type mitochondria treated with 0.8 μ

CsA (○) or CyP-D-null mitochondria (▴) were incubated in the presence of the indicated millimolar concentrations of Pi. B, wild-type (circles) or CyP-D-null (triangles) mitochondria were incubated in the presence of the indicated millimolar concentrations of Vi (open symbols) or Asi (closed symbols). C and D, wild-type mitochondria (•), CyP-D-null mitochondria (▴), or wild-type mitochondria treated with 0.8 μ

CsA (○) were incubated with the indicated millimolar concentrations of Pi and Vi (C) or Pi and Asi (D). The incubation conditions were otherwise identical to those described in the legend to Fig. 1_A_, and the CRC is expressed in nanomoles of Ca2+/mg of protein.

FIGURE 4.

Sensitivity of PTP to inducers is unaffected by CyP-D ablation in the absence of Pi. Wild-type (•) or CyP-D-null (▴) mitochondria were incubated in the presence of 1 m

Vi (A–C) or 1 m

Asi (A_′_–C_′) in the presence of the indicated concentrations of phenylarsine oxide (PhAsO), diamide, or arachidonic acid. Experimental conditions were otherwise identical to those described in the legend to Fig. 1_A, and the CRC is normalized to the value measured in the absence of PTP inducers (CRC0).

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References

1. Kroemer, G., Galluzzi, L., and Brenner, C. (2007) Physiol. Rev. 87 99–163 - PubMed
1. Bernardi, P., Krauskopf, A., Basso, E., Petronilli, V., Blachly-Dyson, E., Di Lisa, F., and Forte, M. A. (2006) FEBS J. 273 2077–2099 - PubMed
1. Fournier, N., Ducet, G., and Crevat, A. (1987) J. Bioenerg. Biomembr. 19 297–303 - PubMed
1. Crompton, M., Ellinger, H., and Costi, A. (1988) Biochem. J. 255 357–360 - PMC - PubMed
1. Broekemeier, K. M., Dempsey, M. E., and Pfeiffer, D. R. (1989) J. Biol. Chem. 264 7826–7830 - PubMed

Phosphate is essential for inhibition of the mitochondrial permeability transition pore by cyclosporin A and by cyclophilin D ablation - PubMed (original) (raw)