Torosyan, H.; Paul, M. D.; Maker, A.; Meyer, B. G.; Jura, N.; Verba, K. A.

PI3K is a potent oncogene that converts PIP2 to PIP3 at the plasma membrane upon activation by receptor tyrosine kinases and Ras GTPases. In the absence of any structures of activated PI3K, the molecular details of its activation remain unknown. Here, we present cryo-EM structures of the PI3K/KRas complex embedded in lipid nanodiscs, revealing a rich ensemble of PI3K states adopted at the membrane surface. The sequential addition of a lipid bilayer, PIP2 and an activating phosphopeptide leads to the progressive release of key inhibitory domains from the PI3K catalytic core, which directly correlates with the reorganization of its active site. While association with POPC/POPS nanodiscs partially relieves PI3K autoinhibition, incorporation of PIP2 triggers near-complete displacement of PI3K inhibitory domains and significant restructuring of active site regulatory motifs. The addition of the activating phosphopeptide induces dimerization of the PI3K/KRas complex through a p110 catalytic subunit-mediated interface that is sterically occluded in autoinhibited PI3K. In cells, this dimeric PI3K complex amplifies Akt signaling in response to growth factor stimulation. Collectively, our structures map the conformational landscape of PI3K activation and reveal previously unexplored interfaces for potential therapeutic targeting.