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Published on: October 20, 2021
Distances and charges along the Orai1 nexus-TM3 interface control STIM1 binding and pore opening
Julia Söllner1, Magdalena Prantl1, Hadil Najjar1
1Institute of Biophysics, JKU Life Science Center, Johannes Kepler University Linz, Gruberstraße 40, 4020 Linz, Austria.
Abstract:
Calcium (Ca2+) influx through the Ca2+ release-activated Ca2+ (CRAC) channel is triggered by binding of the Ca2+ sensor STIM1 to the pore-forming Orai1 complex, primarily to its cytosolic C termini. These C termini connect to transmembrane (TM) domain 4 via the flexible nexus region, which has been proposed to transmit the activation signal toward the central pore via concentrically arranged TM domains. However, the conformational dynamics of the nexus-TM3 interface required for channel gating remain elusive. Here, we combine unnatural amino acid (UAA)-based photocrosslinking and chemical crosslinking with site-directed mutagenesis to investigate this interface. We report that a widening of the nexus-TM3 interface is an essential step within the cascade of conformational rearrangements underlying STIM1-mediated pore opening, while hydrophobicity and charges in this interface are further determinants contributing to signal propagation to the pore. These findings underscore the relevance of the nexus-TM3 dynamics for proper Orai1 function.
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