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纳米抗体与人类酸敏感离子通道1a（ASIC1a）结合的结构与分析

Structure and analysis of nanobody binding to the human ASIC1a ion channel.

作者信息

Wu Yangyu, Chen Zhuyuan, Sigworth Fred J, Canessa Cecilia M

机构信息

Basic Sciences Department, Tsinghua University School of Medicine, Beijing, China.

Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, United States.

出版信息

Elife. 2021 Jul 28;10:e67115. doi: 10.7554/eLife.67115.

DOI:10.7554/eLife.67115
PMID:34319232
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8318589/
Abstract

ASIC1a is a proton-gated sodium channel involved in modulation of pain, fear, addiction, and ischemia-induced neuronal injury. We report isolation and characterization of alpaca-derived nanobodies (Nbs) that specifically target human ASIC1a. Cryo-electron microscopy of the human ASIC1a channel at pH 7.4 in complex with one of these, Nb.C1, yielded a structure at 2.9 Å resolution. It is revealed that Nb.C1 binds to a site overlapping with that of the Texas coral snake toxin (MitTx1) and the black mamba venom Mambalgin-1; however, the Nb.C1-binding site does not overlap with that of the inhibitory tarantula toxin psalmotoxin-1 (PcTx1). Fusion of Nb.C1 with PcTx1 in a single polypeptide markedly enhances the potency of PcTx1, whereas competition of Nb.C1 and MitTx1 for binding reduces channel activation by the toxin. Thus, Nb.C1 is a molecular tool for biochemical and structural studies of hASIC1a; a potential antidote to the pain-inducing component of coral snake bite; and a candidate to potentiate PcTx1-mediated inhibition of hASIC1a in vivo for therapeutic applications.

摘要

ASIC1a是一种质子门控钠通道，参与疼痛、恐惧、成瘾和缺血诱导的神经元损伤的调节。我们报告了从羊驼中分离并鉴定出特异性靶向人ASIC1a的纳米抗体（Nbs）。在pH 7.4条件下，人ASIC1a通道与其中一种纳米抗体Nb.C1形成复合物的冷冻电子显微镜分析得到了分辨率为2.9 Å的结构。结果显示，Nb.C1结合的位点与德州珊瑚蛇毒素（MitTx1）和黑曼巴蛇毒Mambalgin-1的结合位点重叠；然而，Nb.C1的结合位点与抑制性狼蛛毒素Psalmotoxin-1（PcTx1）的结合位点不重叠。在单一多肽中将Nb.C1与PcTx1融合可显著增强PcTx1的效力，而Nb.C1和MitTx1竞争结合会降低毒素对通道的激活作用。因此，Nb.C1是用于hASIC1a生化和结构研究的分子工具；是珊瑚蛇咬伤致痛成分的潜在解毒剂；也是在体内增强PcTx1介导的对hASIC1a抑制作用以用于治疗应用的候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/aca8b825da74/elife-67115-fig5.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/32c43f056dd9/elife-67115-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/aca8b825da74/elife-67115-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/ec6c2407a45f/elife-67115-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/148bff30c4e4/elife-67115-fig1-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/ac0fa1823f1b/elife-67115-fig1-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/f20a880a749e/elife-67115-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/3bd76cd72a05/elife-67115-fig2-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/d7b77f46723b/elife-67115-fig2-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/87095d5186f6/elife-67115-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/200732d301c3/elife-67115-fig3-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/50bf558dfbe4/elife-67115-fig3-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/9eb06890087a/elife-67115-fig3-figsupp3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/33deb243ea66/elife-67115-fig3-figsupp4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/22c48f7697d4/elife-67115-fig3-figsupp5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/32c43f056dd9/elife-67115-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f01/8318589/aca8b825da74/elife-67115-fig5.jpg

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