Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET

Kubota, Tomoya, Durek, Thomas, Dang, Bobo, Finol-Urdaneta, Rocio K., Craik, David J., Kent, Stephen B. H., French, Robert J., Bezanilla, Francisco and Correa, Ana M. (2017) Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET. Proceedings of the National Academy of Sciences, 114 10: E1857-E1865. doi:10.1073/pnas.1700453114


Author Kubota, Tomoya
Durek, Thomas
Dang, Bobo
Finol-Urdaneta, Rocio K.
Craik, David J.
Kent, Stephen B. H.
French, Robert J.
Bezanilla, Francisco
Correa, Ana M.
Title Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET
Journal name Proceedings of the National Academy of Sciences   Check publisher's open access policy
ISSN 1091-6490
0027-8424
Publication date 2017-03-07
Sub-type Article (original research)
DOI 10.1073/pnas.1700453114
Open Access Status Not yet assessed
Volume 114
Issue 10
Start page E1857
End page E1865
Total pages 9
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Language eng
Formatted abstract
Voltage-gated sodium channels (Navs) play crucial roles in excitable cells. Although vertebrate Nav function has been extensively studied, the detailed structural basis for voltage-dependent gating mechanisms remain obscure. We have assessed the structural changes of the Nav voltage sensor domain using lanthanide-based resonance energy transfer (LRET) between the rat skeletal muscle voltage-gated sodium channel (Nav1.4) and fluorescently labeled Nav1.4-targeting toxins. We generated donor constructs with genetically encoded lanthanide-binding tags (LBTs) inserted at the extracellular end of the S4 segment of each domain (with a single LBT per construct). Three different Bodipy-labeled, Nav1.4-targeting toxins were synthesized as acceptors: β-scorpion toxin (Ts1)-Bodipy, KIIIA-Bodipy, and GIIIA-Bodipy analogs. Functional Nav-LBT channels expressed in Xenopus oocytes were voltage-clamped, and distinct LRET signals were obtained in the resting and slow inactivated states. Intramolecular distances computed from the LRET signals define a geometrical map of Nav1.4 with the bound toxins, and reveal voltage-dependent structural changes related to channel gating.
Keyword Relaxed state
Slow inactivation
Voltage gating
β-scorpion toxin
μ-conotoxin
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Institute for Molecular Bioscience - Publications
 
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