Subtype-selective positive modulation of K_Ca 2 channels depends on the HA/HB helices.

ABSTRACT:

Background and purpose

In the activated state of small-conductance Ca²⁺ -activated potassium (K_Ca 2) channels, calmodulin interacts with the HA/HB helices and the S4-S5 linker. CyPPA potentiates K_Ca 2.2a and K_Ca 2.3 channel activity but not the K_Ca 2.1 and K_Ca 3.1 subtypes.

Experimental approach

Site-directed mutagenesis, patch-clamp recordings and in silico modelling were utilised to explore the structural determinants for the subtype-selective modulation of K_Ca 2 channels by CyPPA.

Key results

Mutating residues in the HA (V420) and HB (K467) helices of K_Ca 2.2a channels to their equivalent residues in K_Ca 3.1 channels diminished the potency of CyPPA. CyPPA elicited prominent responses on mutant K_Ca 3.1 channels with an arginine residue in the HB helix substituted for its equivalent lysine residue in the K_Ca 2.2a channels (R355K). K_Ca 2.1 channels harbouring a three-amino-acid insertion upstream of the cognate R438 residues in the HB helix showed no response to CyPPA, whereas the deletion mutant (K_Ca 2.1_ΔA434/Q435/K436) became sensitive to CyPPA. In molecular dynamics simulations, CyPPA docked between calmodulin C-lobe and the HA/HB helices widens the cytoplasmic gate of K_Ca 2.2a channels.

Conclusion and implications

Selectivity of CyPPA among K_Ca 2 and K_Ca 3.1 channel subtypes relies on the HA/HB helices.

SUBMITTER: Nam YW 

PROVIDER: S-EPMC8799485 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature