Open AccessThe distinct role of the four voltage sensors of the skeletal CaV1.1 channel in voltage-dependent activation
Author(s) -
Nicoletta Savalli,
Marina Angelini,
Federica Steccanella,
Julian Wier,
Fuli Wu,
Marbella Quiñonez,
Marino DiFranco,
Alan Neely,
Stephen C. Can,
Riccardo Olcese
Publication year - 2021
Publication title -
the journal of general physiology/the journal of general physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.064
H-Index - 127
eISSN - 1540-7748
pISSN - 0022-1295
DOI - 10.1085/jgp.202112915
Subject(s) - cav1.2 , depolarization , ryr1 , biophysics , chemistry , allosteric regulation , electrophysiology , membrane potential , voltage clamp , gating , intracellular , voltage dependent calcium channel , calcium , biology , ryanodine receptor , neuroscience , biochemistry , enzyme , organic chemistry
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 CaV1.1 channels to inherit voltage sensitivity. Using an opto-electrophysiological approach, we resolved the excitation-driven molecular events controlling both CaV1.1 and RYR1 activations, reported as fluorescence changes. We discovered that each of the four human CaV1.1 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 CaV1.1 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 CaV1.1 activation was also confirmed using a naturally occurring, charge-neutralizing mutation in VSD-I (R174W). This mutation abolished CaV1.1 current at physiological membrane potentials by impairing VSD-I activation without affecting the other VSDs. Using a structurally relevant allosteric model of CaV activation, which accounted for both time- and voltage-dependent properties of CaV1.1, to predict VSD-pore coupling energies, we found that VSD-I contributed the most energy (~75 meV or ∼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 ∼1 kT). This study settles the longstanding question of how CaV1.1, 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 CaV1.1 channels is primarily driven by the activation of one voltage sensor, a mechanism distinct from that of all other voltage-gated channels.