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MAC inhibitors antagonize the pro-apoptotic effects of tBid and disassemble Bax / Bak oligomers

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Abstract

Mitochondrial Apoptotic Channel inhibitors or iMACs are di-bromocarbazole derivatives with anti-apoptotic function which have been tested and validated in several mouse models of brain injury and neurodegeneration. Owing to the increased therapeutic potential of these compounds, we sought to expand our knowledge of their mechanism of action. We investigated the kinetics of MAC inhibition in mitochondria from wild type, Bak, and Bax knockout cell lines using patch clamp electrophysiology, fluorescence microscopy, ELISA, and semiquantitative western blot analyses. Our results show that iMACs work through at least two mechanisms: 1) by blocking relocation of the cytoplasmic Bax protein to mitochondria and 2) by disassembling Bax and Bak oligomers in the mitochondrial outer membrane. iMACs exert comparable effects on channel conductance of Bax or Bak and similarly affect cytochrome c release from Bax or Bak-containing mitochondria. Interestingly, wild type mitochondria were more susceptible to inhibition than the Bak or Bax knockouts. Western blot analysis showed that wild type mitochondria had lower steady state levels of Bak in the absence of apoptotic stimulation.

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Acknowledgments

This study was funded by CUNY/CIRG-2265 award to PMP, NIH award GM57249 to KK. We thank David Andrews (University of Toronto) for providing Smac-Cherry plasmids and Gorka Basañez (University of Basque Country) for the recombinant Bak protein.

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Authors and Affiliations

  1. Baruch College and Graduate Center of City University of New York (CUNY), New York, NY, 10010, USA

    Pablo M. Peixoto & Oygul Mirzalieva

  2. Department of Chemistry, California State University of Fresno, Fresno, CA, 93740, USA

    Laurent M. Dejean

  3. New York University College of Dentistry, New York, NY, 10010, USA

    Pablo M. Peixoto, Evgeny V. Pavlov & Kathleen W. Kinnally

  4. Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA

    Oscar Teijido

  5. Merck Serono, Geneva Research Center, Geneva, 1202, Switzerland

    Bruno Antonsson

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  1. Pablo M. Peixoto
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  2. Oscar Teijido
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  3. Oygul Mirzalieva
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  4. Laurent M. Dejean
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Correspondence to Pablo M. Peixoto.

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Peixoto, P.M., Teijido, O., Mirzalieva, O. et al. MAC inhibitors antagonize the pro-apoptotic effects of tBid and disassemble Bax / Bak oligomers. J Bioenerg Biomembr 49, 65–74 (2017). https://doi.org/10.1007/s10863-015-9635-7

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  • DOI: https://doi.org/10.1007/s10863-015-9635-7

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