A tarantula-venom peptide antagonizes the TRPA1 nociceptor ion channel by binding to the S1-S4 gating domain.
A tarantula-venom peptide antagonizes the TRPA1 nociceptor ion channel by binding to the S1-S4 gating domain.
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DOI：
10.1016/j.cub.2014.01.013
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发表时间：
2014-03-03
期刊：
影响因子：
9.2
通讯作者：
Nitabach, Michael N.
中科院分区：
文献类型：
--
作者：
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.
蜘蛛、蝎子、锥螺、海葵和蛇等食肉动物的毒液是药物发现的药理学多样性的极好来源，也是阐明离子通道结构、功能和生理特性的药理学工具。在这里，我们描述了第一个已知的伤害感受器离子通道瞬时受体电位锚蛋白1（TRPA 1）的肽拮抗剂。我们构建了一个重组cDNA文库，编码1000多种GPI锚定肽毒素（T毒素）来自蜘蛛毒液，并筛选该文库在非洲爪蟾卵母细胞与TRPA 1共表达。该筛选导致原毒素-I（ProTx-I）的鉴定，其是来自秘鲁绿绒狼蛛Thrixopelma pruriens的毒液的35个残基的肽，作为第一个已知的高亲和力肽TRPA 1拮抗剂。有趣的是，ProTx-I之前被确定为电压门控钠（NaV）通道的拮抗剂。为了鉴定ProTx-I与这些不同离子通道类型结合的表面，我们构建了ProTx-I丙氨酸扫描突变体的t毒素文库，并针对NaV1.2和TRPA 1筛选该文库。这揭示了ProTx-I与这两种离子通道结合的不同部分重叠表面，其特定的化学特征解释了其对NaV1.2的亲和力高于TRPA 1。重要的是，这种诱变产生了两种新的ProTx-I变体，它们仅对TRPA 1或NaV 1.2有活性，但不能同时对两者都有活性。通过测试其对嵌合通道的活性，我们将TRPA 1 S1-S4门控结构域的细胞外环鉴定为ProTx-I结合位点。这些研究建立了筛选天然和突变毒素的t-毒素库，我们称之为“毒素库”，作为分离新型离子通道修饰剂和设计具有改变的靶选择性的离子通道修饰剂的普遍适用的方法。他们还表明，ProTx-I将是一种有价值的药理学试剂，用于解决TRPA 1门控的生物物理机制，TRPA 1在伤害感受器中功能的生理学和病理生理学，以及在疼痛和炎症背景下的潜在临床应用。
The venoms of predators such as spiders, scorpions, cone snails, sea anemones, and snakes, have been an excellent source of pharmacological diversity for drug discovery and as pharmacological tools for elucidating the structure, function, and physiological properties of ion channels. Here we describe the first known peptide antagonist of the nociceptor ion channel transient receptor potential ankyrin 1 (TRPA1). We constructed a recombinant cDNA library encoding ∼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. Interestingly, ProTx-I was previously identified as an antagonist of voltage-gated sodium (NaV) channels. To identify the surfaces of ProTx-I by which it binds to these distinct ion channel types, 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, and whose specific chemical features explain its higher affinity for NaV1.2 than for TRPA1. Importantly, this mutagenesis yielded two novel ProTx-I variants that are only active against either TRPA1or NaV1.2, but not both. 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. These studies establish screening of t-toxin libraries of native and mutated toxins, which we term “toxineering”, as a generally applicable method for isolation of novel ion channel modifiers and for design of ion channel modifiers with altered target selectivity. They also suggest that ProTx-I will be a valuable pharmacological reagent for addressing the biophysical mechanisms of TRPA1 gating, the physiology and pathophysiology of TRPA1 function in nociceptors, and for potential clinical application in the context of pain and inflammation.
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DOI：
10.1523/jneurosci.5369-07.2008
发表时间：
2008-03-05
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Andersson DA;Gentry C;Moss S;Bevan S
通讯作者：
Bevan S
DOI：
10.1073/pnas.0812675106
发表时间：
2009-05-19
影响因子：
11.1
作者：
Andersson, David A.;Gentry, Clive;Bevan, Stuart
通讯作者：
Bevan, Stuart
影响因子：
15.9
作者：
Dai, Yi;Wang, Shenglan;Noguchi, Koichi
通讯作者：
Noguchi, Koichi
狼蛛Chilobrachys jingzhao胱氨酸结毒素的分子多样性和进化
影响因子：
8
作者：
Chen, J.;Deng, M.;Liang, S.
通讯作者：
Liang, S.
影响因子：
15.9
作者：
Bessac, Bret F.;Sivula, Michael;Jordt, Sven-Eric
通讯作者：
Jordt, Sven-Eric