Bcl-2 Oligomerizes Bax on the Mitochondrial Membrane Surface Preventing the Initial Stages of Apoptosis

Ayscough, S. E.; Clifton, L. A.; Aden, J.; Köhler, S.; Paracini, N.; Doutch, J.; Bragginton, E. C.; Leung, A. E.; Bogojevic, O.; Poon, J.-F.; Nagy, T. M.; Wacklin-Knecht, H. P.; Gröbner, G.

doi:10.1101/2025.01.24.634681

bioRxiv · 10.1101/2025.01.24.634681

Bcl-2 Oligomerizes Bax on the Mitochondrial Membrane Surface Preventing the Initial Stages of Apoptosis

Ayscough, S. E.·Clifton, L. A.·Aden, J.·Köhler, S.·Paracini, N.·Doutch, J.·Bragginton, E. C.·Leung, A. E.·Bogojevic, O.·Poon, J.-F.·Nagy, T. M.·Wacklin-Knecht, H. P.·Gröbner, G.

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Abstract

The Bcl-2 family of proteins control mitochondrial outer membrane (MOM) pore formation, crucial to cellular clearance via apoptosis. However, the molecular principles by which opposing family members inhibit, mediate or promote MOM perforation remain elusive. Here, we demonstrate that cell-protecting Bcl-2 directly sequesters cell-killing Bax into a protein-protein complex at the membrane interface preventing Bax forming apoptotic pores. Neutron reflectometry showed Bax association with Bcl-2 occurs through the formation of Bax protein oligomers on the membrane surface. Bax binding to the membrane surface was proportional to the membrane-embedded Bcl-2 suggestive of protein-protein complex formation through both Bcl-2/Bax and Bax/Bax interactions. Bcl-2/Bax sequestration to prevent perforation was observed in membrane models with and without pro-apoptotic cardiolipin present. Our findings shed fundamental new light on the communication of Bcl-2 with its cell-killing relatives at the mitochondrias membraneous exterior to prevent cells from undergoing apoptosis. TeaserMembrane-embedded Bcl-2 sequesters Bax, to prevent the perforation of mitochondrial membranes by Bax which would initiate apoptosis.

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Ayscough, S. E., Clifton, L. A., Aden, J., Köhler, S., Paracini, N., Doutch, J., Bragginton, E. C., Leung, A. E., Bogojevic, O., Poon, J.-F., Nagy, T. M., Wacklin-Knecht, H. P., Gröbner, G.. 2025-01-27. Bcl-2 Oligomerizes Bax on the Mitochondrial Membrane Surface Preventing the Initial Stages of Apoptosis. https://doi.org/10.1101/2025.01.24.634681

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