Protein–Lipid Interfaces Can Drive the Functions of Membrane-Embedded Protein–Protein Complexes

[Image: see text] The roles of surrounding membrane lipids in the functions of transmembrane and peripheral membrane proteins are largely unknown. Herein, we utilize the recently reported structures of the TRPV1 ion channel protein bound to its potent protein agonist, the double-knot toxin (DkTx), as a model system to investigate the roles of toxin–lipid interfaces in TRPV1 activation by characterizing a series of DkTx variants electrophysiologically. Together with membrane partitioning experiments, these studies reveal that toxin–lipid interfaces play an overwhelmingly dominant role in channel activation as compared to lipid-devoid toxin–channel interfaces. Additionally, we find that whereas the membrane interfaces formed by one of the knots of the toxin endow it with its low channel-dissociation rate, those formed by other knot contribute primarily to its potency. These studies establish that protein–lipid interfaces play nuanced yet profound roles in the function of protein–protein complexes within membranes.