Regulation of Kv2.1 channel inactivation by phosphatidylinositol 4,5-bisphosphate

Phosphatidylinositol 4,5-bisphosphate (PIP₂) is a membrane phospholipid that regulates the function of multiple ion channels, including some members of the voltage-gated potassium (Kv) channel superfamily. The PIP₂ sensitivity of Kv channels is well established for all five members of the Kv7 family and for Kv1.2 channels; however, regulation of other Kv channels by PIP₂ remains unclear. Here, we investigate the effects of PIP₂ on Kv2.1 channels by applying exogenous PIP₂ to the cytoplasmic face of excised membrane patches, activating muscarinic receptors (M1R), or depleting endogenous PIP₂ using a rapamycin-translocated 5-phosphatase (FKBP-Inp54p). Exogenous PIP₂ rescued Kv2.1 channels from rundown and partially prevented the shift in the voltage-dependence of inactivation observed in inside-out patch recordings. Native PIP₂ depletion by the recruitment of FKBP-Insp54P or M1R activation in whole-cell experiments, induced a shift in the voltage-dependence of inactivation, an acceleration of the closed-state inactivation, and a delayed recovery of channels from inactivation. No significant effects were observed on the activation mechanism by any of these treatments. Our data can be modeled by a 13-state allosteric model that takes into account that PIP₂ depletion facilitates inactivation of Kv2.1. We propose that PIP₂ regulates Kv2.1 channels by interfering with the inactivation mechanism.