Raw data for PIP2 interaction with TRPC3, explored through computation and electrophysiology

The transient receptor potential canonical type 3 (TRPC3) channel plays a pivotal role in regulating neuronal excitability within the brain via its constitutive activity. The channel is intricately regulated by lipids and has previously been demonstrated to be positively modulated by PIP2. Using molecular dynamics simulations and patch clamp techniques, we reveal that PIP2 predominantly interacts with TRPC3 at the L3 lipid binding site, located at the intersection of pre-S1 and S1 helices. We propose a novel signal transduction pathway from the L3 through the re-entrant loop to a salt bridge between the TRP helix and S4-S5 linker. Notably, we find that both stimulated and constitutive TRPC3 activity require PIP2. These structural insights into the function of TRPC3 are invaluable for understanding the role of the TRPC subfamily in health and disease in native tissue.