Molecular Determinant for Specific Ca/Ba Selectivity Profiles of Low and High Threshold Ca²⁺ Channels

Voltage-gated Ca²⁺ channels (VGCC) play a key role in many physiological functions by their high selectivity for Ca²⁺ over other divalent and monovalent cations in physiological situations. Divalent/monovalent selection is shared by all VGCC and is satisfactorily explained by the existence, within the pore, of a set of four conserved glutamate/aspartate residues (EEEE locus) coordinating Ca²⁺ ions. This locus however does not explain either the choice of Ca²⁺ among other divalent cations or the specific conductances encountered in the different VGCC. Our systematic analysis of high- and low-threshold VGCC currents in the presence of Ca²⁺ and Ba²⁺ reveals highly specific selectivity profiles. Sequence analysis, molecular modeling, and mutational studies identify a set of nonconserved charged residues responsible for these profiles. In HVA (high voltage activated) channels, mutations of this set modify divalent cation selectivity and channel conductance without change in divalent/monovalent selection, activation, inactivation, and kinetics properties. The Ca_V2.1 selectivity profile is transferred to Ca_V2.3 when exchanging their residues at this location. Numerical simulations suggest modification in an external Ca²⁺ binding site in the channel pore directly involved in the choice of Ca²⁺, among other divalent physiological cations, as the main permeant cation for VGCC. In LVA (low voltage activated) channels, this locus (called DCS for divalent cation selectivity) also influences divalent cation selection, but our results suggest the existence of additional determinants to fully recapitulate all the differences encountered among LVA channels. These data therefore attribute to the DCS a unique role in the specific shaping of the Ca²⁺ influx between the different HVA channels.