Unknown

Dataset Information

0

Mutations of nonconserved residues within the calcium channel alpha1-interaction domain inhibit beta-subunit potentiation.


ABSTRACT: Voltage-dependent calcium channels consist of a pore-forming subunit (Ca(V)alpha(1)) that includes all the molecular determinants of a voltage-gated channel, and several accessory subunits. The ancillary beta-subunit (Ca(V)beta) is a potent activator of voltage-dependent calcium channels, but the mechanisms and structural bases of this regulation remain elusive. Ca(V)beta binds reversibly to a conserved consensus sequence in Ca(V)alpha(1), the alpha(1)-interaction domain (AID), which forms an alpha-helix when complexed with Ca(V)beta. Conserved aromatic residues face to one side of the helix and strongly interact with a hydrophobic pocket on Ca(V)beta. Here, we studied the effect of mutating residues located opposite to the AID-Ca(V)beta contact surface in Ca(V)1.2. Substitution of AID-exposed residues by the corresponding amino acids present in other Ca(V)alpha(1) subunits (E462R, K465N, D469S, and Q473K) hinders Ca(V)beta's ability to increase ionic-current to charge-movement ratio (I/Q) without changing the apparent affinity for Ca(V)beta. At the single channel level, these Ca(V)1.2 mutants coexpressed with Ca(V)beta(2a) visit high open probability mode less frequently than wild-type channels. On the other hand, Ca(V)1.2 carrying either a mutation in the conserved tryptophan residue (W470S, which impairs Ca(V)beta binding), or a deletion of the whole AID sequence, does not exhibit Ca(V)beta-induced increase in I/Q. In addition, we observed a shift in the voltage dependence of activation by +12 mV in the AID-deleted channel in the absence of Ca(V)beta, suggesting a direct participation of these residues in the modulation of channel activation. Our results show that Ca(V)beta-dependent potentiation arises primarily from changes in the modal gating behavior. We envision that Ca(V)beta spatially reorients AID residues that influence the channel gate. These findings provide a new framework for understanding modulation of VDCC gating by Ca(V)beta.

SUBMITTER: Gonzalez-Gutierrez G 

PROVIDER: S-EPMC2518731 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature

altmetric image

Publications

Mutations of nonconserved residues within the calcium channel alpha1-interaction domain inhibit beta-subunit potentiation.

Gonzalez-Gutierrez Giovanni G   Miranda-Laferte Erick E   Naranjo David D   Hidalgo Patricia P   Neely Alan A  

The Journal of general physiology 20080901 3


Voltage-dependent calcium channels consist of a pore-forming subunit (Ca(V)alpha(1)) that includes all the molecular determinants of a voltage-gated channel, and several accessory subunits. The ancillary beta-subunit (Ca(V)beta) is a potent activator of voltage-dependent calcium channels, but the mechanisms and structural bases of this regulation remain elusive. Ca(V)beta binds reversibly to a conserved consensus sequence in Ca(V)alpha(1), the alpha(1)-interaction domain (AID), which forms an al  ...[more]

Similar Datasets

| S-EPMC1563498 | biostudies-literature
| S-EPMC2287295 | biostudies-literature
| S-EPMC6368824 | biostudies-literature
| S-EPMC5549883 | biostudies-literature
| S-EPMC6579206 | biostudies-literature
| S-EPMC3076333 | biostudies-literature
| S-EPMC1275639 | biostudies-literature
| S-EPMC6758875 | biostudies-literature
2023-09-27 | GSE224912 | GEO
| S-EPMC23172 | biostudies-literature