Molecular basis of the PIP2-dependent regulation of Ca(v)2.2 channel and its modulation by Ca-v beta subunits

Abstract

High-voltage-activated Ca2+ (CaV) channels that adjust Ca2+ influx upon membrane depolarization are differentially regulated by phosphatidylinositol 4,5-bisphosphate (PIP2) in an auxiliary CaV β subunit-dependent manner. However, the molecular mechanism by which the β subunits control the PIP2 sensitivity of CaV channels remains unclear. By engineering various α1B and β constructs in tsA-201 cells, we reported that at least two PIP2-binding sites, including the polybasic residues at the C-terminal end of I–II loop and the binding pocket in S4II domain, exist in the CaV2.2 channels. Moreover, they were distinctly engaged in the regulation of channel gating depending on the coupled CaV β2 subunits. The membrane-anchored β subunit abolished the PIP2 interaction of the phospholipid-binding site in the I–II loop, leading to lower PIP2 sensitivity of CaV2.2 channels. By contrast, PIP2 interacted with the basic residues in the S4II domain of CaV2.2 channels regardless of β2 isotype. Our data demonstrated that the anchoring properties of CaV β2 subunits to the plasma membrane determine the biophysical states of CaV2.2 channels by regulating PIP2 coupling to the nonspecific phospholipid-binding site in the I–II loop. © 2022, eLife Sciences Publications Ltd. All rights reserved.