The distinct role of the four voltage sensors of the skeletal Ca(V)1.1 channel in voltage-dependent activation
Initiation of skeletal muscle contraction is triggered by rapid activation of RYR1 channels in response to sarcolemmal depolarization. RYR1 is intracellular and has no voltage-sensing structures, but it is coupled with the voltage-sensing apparatus of Ca(V)1.1 channels to inherit voltage sensitivity. Using an opto-electrophysiological approach, we resolved the excitation-driven molecular events controlling both Ca(V)1.1 and RYR1 activations, reported as fluorescence changes. We discovered that each of the four human Ca(V)1.1 voltage-sensing domains (VSDs) exhibits unique biophysical properties: VSD-I time-dependent properties were similar to ionic current activation kinetics, suggesting a critical role of this voltage sensor in Ca(V)1.1 activation; VSD-II, VSD-III, and VSD-IV displayed faster activation, compatible with kinetics of sarcoplasmic reticulum Ca2+ release. The prominent role of VSD-I in governing Ca(V)1.1 activation was also confirmed using a naturally occurring, chargeneutralizing mutation in VSD-I (R174W). This mutation abolished Ca(V)1.1 current at physiological membrane potentials by impairing VSD-I activation without affecting the other VSDs. Using a structurally relevant allosteric model of Ca-V activation, which accounted for both time- and voltage-dependent properties of Ca(V)1.1, to predict VSD-pore coupling energies, we found that VSD-I contributed the most energy (similar to 75 meV or similar to 3 kT) toward the stabilization of the open states of the channel, with smaller (VSD-IV) or negligible (VSDs II and III) energetic contribution from the other voltage sensors (<25 meV or similar to 1 kT). This study settles the longstanding question of how Ca(V)1.1, a slowly activating channel, can trigger RYR1 rapid activation, and reveals a new mechanism for voltage-dependent activation in ion channels, whereby pore opening of human Ca(V)1.1 channels is primarily driven by the activation of one voltage sensor, a mechanism distinct from that of all other voltage-gated channels.
|Título según WOS:||The distinct role of the four voltage sensors of the skeletal Ca(V)1.1 channel in voltage-dependent activation|
|Título de la Revista:||JOURNAL OF GENERAL PHYSIOLOGY|
|Editorial:||ROCKEFELLER UNIV PRESS|
|Fecha de publicación:||2021|