Sites Contributing to TRPA1 Activation by the Anesthetic Propofol Identified by Photoaffinity Labeling
Access Status
Full text of the requested work is not available in DASH at this time ("restricted access"). For more information on restricted deposits, see our FAQ.Author
Woll, Kellie A.
Gianti, Eleonora
Bhanu, Natarajan V.
Garcia, Benjamin A.
Carnevale, Vincenzo
Eckenhoff, Roderic G.
Published Version
https://doi.org/10.1016/j.bpj.2017.08.040Metadata
Show full item recordCitation
Woll, Kellie A., Kenneth A. Skinner, Eleonora Gianti, Natarajan V. Bhanu, Benjamin A. Garcia, Vincenzo Carnevale, Roderic G. Eckenhoff, and Rachelle Gaudet. 2017. “Sites Contributing to TRPA1 Activation by the Anesthetic Propofol Identified by Photoaffinity Labeling.” Biophysical Journal 113 (10) (November): 2168–2172. doi:10.1016/j.bpj.2017.08.040.Abstract
In addition to inducing anesthesia, propofol activates a key component of the pain pathway, the transient receptor potential ankyrin 1 ion channel (TRPA1). Recent mutagenesis studies suggested a potential activation site within the transmembrane domain, near the A-967079 cavity. However, mutagenesis cannot distinguish between protein-based and ligand-based mechanisms, nor can this site explain the complex modulation by propofol. Thus more direct approaches are required to reveal potentially druggable binding sites. Here we apply photoaffinity labeling using a propofol derivative, meta-azipropofol, for direct identification of binding sites in mouse TRPA1. We confirm that meta-azipropofol activates TRPA1 like the parent anesthetic, and identify two photolabeled residues (V954 and E969) in the S6 helix. In combination with docking to closed and open state models of TRPA1, photoaffinity labeling suggested that the A-967079 cavity is a positive modulatory site for propofol. Further, the photoaffinity labeling of E969 indicated pore block as a likely mechanism for propofol inhibition at high concentrations. The direct identification of drug-binding sites clarifies the molecular mechanisms of important TRPA1 agonists, and will facilitate drug design efforts to modulate TRPA1.Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:34864120
Collections
- FAS Scholarly Articles [18276]
Contact administrator regarding this item (to report mistakes or request changes)