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Synthetic ion transporters can induce apoptosis by facilitating chloride anion transport into cells

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Abstract

Anion transporters based on small molecules have received attention as therapeutic agents because of their potential to disrupt cellular ion homeostasis. However, a direct correlation between a change in cellular chloride anion concentration and cytotoxicity has not been established for synthetic ion carriers. Here we show that two pyridine diamide-strapped calix[4]pyrroles induce coupled chloride anion and sodium cation transport in both liposomal models and cells, and promote cell death by increasing intracellular chloride and sodium ion concentrations. Removing either ion from the extracellular media or blocking natural sodium channels with amiloride prevents this effect. Cell experiments show that the ion transporters induce the sodium chloride influx, which leads to an increased concentration of reactive oxygen species, release of cytochrome c from the mitochondria and apoptosis via caspase activation. However, they do not activate the caspase-independent apoptotic pathway associated with the apoptosis-inducing factor. Ion transporters, therefore, represent an attractive approach for regulating cellular processes that are normally controlled tightly by homeostasis.

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Figure 1: Structures of compounds.
Figure 2: Transport studies using liposomal model membranes.
Figure 3: Ion transporters induce apoptosis.
Figure 4: Ion transporters induce caspase activation but do not activate the AIF-associated apoptotic pathway.
Figure 5: Effect of ions on transporter-induced cell death.

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  • 05 September 2014

    The authors wish to add a sentence to the Acknowledgements: "A part of this work was carried out with support from the Chemical Biology Research Center in Korea Research Institute of Bioscience and Biotechnology." This has been added in all versions of the Article.

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Acknowledgements

This work was supported by the National Creative Research Initiative (grant no. 2010-0018272 to I.S.) program in Korea, as well as by the Office of Basic Energy Sciences, US Department of Energy (grant no. DE-FG02-01ER15186 to J.L.S.). P.A.G. thanks the Engineering and Physical Sciences Research Council for a postdoctoral fellowship (N.B.) (EP/J009687/1). W.VR. and P.A.G. thank the European Union for a Marie Curie Career Integration grant. A part of this work was carried out with support from the Chemical Biology Research Center in Korea Research Institute of Bioscience and Biotechnology.

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Authors and Affiliations

Authors

Contributions

J.L.S, I.S. and P.A.G. designed the study and supervised the work. S-K.K. performed biological studies. S.K.K. designed and synthesized compounds and performed ion-binding studies in solution. S.K.K. and V.M.L. carried out the X-ray single-crystal structure analysis. P.A.G., W.VR., N.B. and A.S. designed and performed the ion-transport studies in liposomes. J.P. and W.N. carried out ion-transport activity studies in cells.

Corresponding authors

Correspondence to Philip A. Gale, Jonathan L. Sessler or Injae Shin.

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The authors declare no competing financial interests.

Supplementary information

Supplementary information

Supplementary information (PDF 10471 kb)

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Crystallographic data for 1·CsCl (CIF 1298 kb)

Supplementary information

Crystallographic data for 12·NaCl·TMACl·(CH3OH)2·(H2O)2 (CIF 1601 kb)

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Crystallographic data for 2·CsCl (CIF 1206 kb)

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Crystallographic data for 22·NaCl·TMACl·(H2O)5·C6H14 (CIF 2987 kb)

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Crystallographic data for 2·TEACl (CIF 39 kb)

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Crystallographic data for 1·(CH3OH)2 (CIF 29 kb)

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Crystallographic data for 3·(CH3OH)2 (CIF 29 kb)

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Ko, SK., Kim, S., Share, A. et al. Synthetic ion transporters can induce apoptosis by facilitating chloride anion transport into cells. Nature Chem 6, 885–892 (2014). https://doi.org/10.1038/nchem.2021

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