MAC inhibitors antagonize the pro-apoptotic effects of tBid and disassemble Bax / Bak oligomers
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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.
KeywordsMAC inhibitors Bax Bak tBid Apoptosis MOMP Patch clamp
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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Conflict of interest
The authors declare no conflict of interest.
- Ding J, Mooers BH, Zhang Z, Kale J, Falcone D, McNichol J et al (2014) After embedding in membranes antiapoptotic Bcl-XL protein binds both Bcl-2 homology region 3 and helix 1 of proapoptotic Bax protein to inhibit apoptotic mitochondrial permeabilization. J Biol Chem 289(17):11873–11896. doi: 10.1074/jbc.M114.552562 CrossRefGoogle Scholar
- Naidoo J, De Jesus-Cortes H, Huntington P, Estill S, Morlock LK, Starwalt R et al (2014) Discovery of a neuroprotective chemical, (S)-N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-fluoropropyl)-6-methoxypyridin-2-amine [(−)-P7C3-S243], with improved druglike properties. J Med Chem 57(9):3746–3754. doi: 10.1021/jm401919s CrossRefGoogle Scholar