期刊
CURRENT BIOLOGY
卷 24, 期 5, 页码 473-483出版社
CELL PRESS
DOI: 10.1016/j.cub.2014.01.013
关键词
-
资金
- NIH [R01NS055035, R01NS056443, R21NS058330, R01GM098931, R01ES015056, U01ES015674]
- Australian Research Council [DP1095728]
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 similar to 100 diverse GPI-anchored peptide toxins (t-toxins) derived from spider venoms and screened this library by co-expression 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 (Na-V) channels. We constructed a t-toxin library of ProTx-I alanine-scanning mutants and screened this library against Na(V)1.2 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 TRPA1 or Na(V)1.2. 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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据