Structure and analysis of nanobody binding to the human ASIC1a ion channel
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-07-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/67115 |
| _version_ | 1811180338559320064 |
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| author | Yangyu Wu Zhuyuan Chen Fred J Sigworth Cecilia M Canessa |
| author_facet | Yangyu Wu Zhuyuan Chen Fred J Sigworth Cecilia M Canessa |
| author_sort | Yangyu Wu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:01:28Z |
| format | Article |
| id | doaj.art-3015e42f8a4b4c0d9076d62eb3fb7ffc |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:01:28Z |
| publishDate | 2021-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3015e42f8a4b4c0d9076d62eb3fb7ffc2022-12-22T04:32:46ZengeLife Sciences Publications LtdeLife2050-084X2021-07-011010.7554/eLife.67115Structure and analysis of nanobody binding to the human ASIC1a ion channelYangyu Wu0https://orcid.org/0000-0001-8064-6132Zhuyuan Chen1Fred J Sigworth2https://orcid.org/0000-0002-7178-8494Cecilia M Canessa3https://orcid.org/0000-0001-7316-5082Basic Sciences Department, Tsinghua University School of Medicine, Beijing, ChinaBasic Sciences Department, Tsinghua University School of Medicine, Beijing, ChinaCellular and Molecular Physiology, Yale University School of Medicine, New Haven, United StatesBasic Sciences Department, Tsinghua University School of Medicine, Beijing, China; Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, United StatesASIC1a 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.https://elifesciences.org/articles/67115ASIC1nanobodycryo-electron microscopyPctx1 potentiationMitTx antagonism |
| spellingShingle | Yangyu Wu Zhuyuan Chen Fred J Sigworth Cecilia M Canessa Structure and analysis of nanobody binding to the human ASIC1a ion channel eLife ASIC1 nanobody cryo-electron microscopy Pctx1 potentiation MitTx antagonism |
| title | Structure and analysis of nanobody binding to the human ASIC1a ion channel |
| title_full | Structure and analysis of nanobody binding to the human ASIC1a ion channel |
| title_fullStr | Structure and analysis of nanobody binding to the human ASIC1a ion channel |
| title_full_unstemmed | Structure and analysis of nanobody binding to the human ASIC1a ion channel |
| title_short | Structure and analysis of nanobody binding to the human ASIC1a ion channel |
| title_sort | structure and analysis of nanobody binding to the human asic1a ion channel |
| topic | ASIC1 nanobody cryo-electron microscopy Pctx1 potentiation MitTx antagonism |
| url | https://elifesciences.org/articles/67115 |
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