Lafargue, E.; Duneau, J.-P.; Buzhinsky, N.; Ornelas, P.; Ortega, A.; Ravishankar, V.; Sturgis, J. N.; Casuso, I.; Bergdoll, l.

Mitochondrial physiology is intricately linked to the oligomerization of the voltage-dependent anion channel (VDAC), acting as the gatekeeper of mitochondria. However, the determinants of VDAC oligomerization remain poorly understood. Here, we used atomic force microscopy to investigate the effects of three primary lipids of the mitochondrial outer membrane (MOM) on VDAC assemblies. We observed that VDAC forms lipid-sensitive clusters termed honeycombs, with their compaction regulated by cholesterol. Molecular dynamics simulations revealed VDAC\'s higher affinity for cholesterol and phosphatidylethanolamine, both impacting the formation of these honeycombs. While we identified honeycomb-like assemblies akin to those in the native MOM, deviations from the physiological lipid composition resulted in varying degrees of disruption of these native-like structures. This emphasizes the profound impact of the lipid environment on VDAC organization. These findings underscore the physiological significance of lipid heterogeneity and changes within biological membranes arising from membrane contacts or pathologies in modulating VDAC behavior.