A tarantula-venom peptide antagonizes the TRPA1 nociceptor ion channel by binding to the S1–S4 gating domain

Gui, Junhong, Liu, Boyi, Cao, Guan, Lipchik, Andrew M., Perez, Minervo, Dekan, Zoltan, Mobli, Mehdi, Daly, Norelle L., Alewood, Paul F., Parker, Laurie L., King, Glenn F., Zhou, Yufeng, Jordt, Sven-Eric and Nitabach, Michael N. (2014) A tarantula-venom peptide antagonizes the TRPA1 nociceptor ion channel by binding to the S1–S4 gating domain. Current Biology, 24 5: 473-483. doi:10.1016/j.cub.2014.01.013

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Author Gui, Junhong
Liu, Boyi
Cao, Guan
Lipchik, Andrew M.
Perez, Minervo
Dekan, Zoltan
Mobli, Mehdi
Daly, Norelle L.
Alewood, Paul F.
Parker, Laurie L.
King, Glenn F.
Zhou, Yufeng
Jordt, Sven-Eric
Nitabach, Michael N.
Title A tarantula-venom peptide antagonizes the TRPA1 nociceptor ion channel by binding to the S1–S4 gating domain
Journal name Current Biology   Check publisher's open access policy
ISSN 0960-9822
Publication date 2014-02-13
Sub-type Article (original research)
DOI 10.1016/j.cub.2014.01.013
Open Access Status DOI
Volume 24
Issue 5
Start page 473
End page 483
Total pages 11
Place of publication Cambridge, MA, United States
Publisher Cell Press
Language eng
Formatted abstract
• The TRPA1 antagonist ProTx-I is identified by screening library of spider toxins
• ProTx-I inhibits NaV channel by stabilizing closed state of voltage sensor
• ProTx-I binds to TRPA1 and NaV by overlapping pharmacophore surfaces
• The S1–S2 extracellular loop participates in ProTx-I binding to TRPA1

Background The venoms of predators have been an excellent source of diverse highly specific peptides targeting ion channels. Here we describe the first known peptide antagonist of the nociceptor ion channel transient receptor potential ankyrin 1 (TRPA1).

Results We constructed a recombinant cDNA library encoding ~100 diverse GPI-anchored peptide toxins (t-toxins) derived from spider venoms and screened this library by coexpression in Xenopus oocytes with TRPA1. This screen resulted in identification of protoxin-I (ProTx-I), a 35-residue peptide from the venom of the Peruvian green-velvet tarantula, Thrixopelma pruriens, as the first known high-affinity peptide TRPA1 antagonist. ProTx-I was previously identified as an antagonist of voltage-gated sodium (NaV) channels. We constructed a t-toxin library of ProTx-I alanine-scanning mutants and screened this library against NaV1.2 and TRPA1. This revealed distinct partially overlapping surfaces of ProTx-I by which it binds to these two ion channels. Importantly, this mutagenesis yielded two novel ProTx-I variants that are only active against either TRPA1or NaV1.2. By testing its activity against chimeric channels, we identified the extracellular loops of the TRPA1 S1–S4 gating domain as the ProTx-I binding site.

Conclusions These studies establish our approach, which we term “toxineering,” as a generally applicable method for isolation of novel ion channel modifiers and design of ion channel modifiers with altered specificity. They also suggest that ProTx-I will be a valuable pharmacological reagent for addressing biophysical mechanisms of TRPA1 gating and the physiology of TRPA1 function in nociceptors, as well as for potential clinical application in the context of pain and inflammation.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Available online 13 February 2014

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
Institute for Molecular Bioscience - Publications
Centre for Advanced Imaging Publications
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Citation counts: TR Web of Science Citation Count  Cited 21 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 25 times in Scopus Article | Citations
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Created: Thu, 20 Feb 2014, 00:55:20 EST by Sandrine Ducrot on behalf of Institute for Molecular Bioscience