An experimental test of the nicotinic hypothesis of COVID-19.
An experimental test of the nicotinic hypothesis of COVID-19.
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DOI：
10.1073/pnas.2204242119
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发表时间：
2022-11
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
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作者：
尽管对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）已经了解了很多，但迄今为止，急性和急性后（“长期”）COVID-19的大量不明症状值得寻找其他相关的生化途径。在氨基酸序列分析和计算方法的基础上，最近有人提出，烟碱乙酰胆碱受体（AChRs）可能作为额外的SARS-CoV-2质膜受体，通过结合病毒的刺突蛋白与正构，乙酰胆碱结合位点重叠的网站。在此，基于实验性配体结合竞争分析，我们得出结论，SARS-CoV-2和胆碱能配体与人α7-AChR的相互排斥结合不太可能是这种复杂疾病的相关方面。COVID-19特征性症状群背后的病理生理机制尚不完全清楚。为了填补这些空白，最近提出了一种“烟碱假说”，即烟碱乙酰胆碱受体（AChR）作为额外的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）受体。该提议的一个关键特征（具有潜在的临床影响）是病毒刺突蛋白和小分子胆碱能配体之间对受体正构结合位点的竞争。这一概念让人想起狂犬病病毒感染期间肌肉AChR的既定作用。为了直接解决这一假设，我们使用同源的人α7-AChR（在完整细胞上表达）作为受体，放射性标记的α-银环蛇毒素（α-BgTx）作为正构位点竞争配体，进行了平衡型配体结合竞争试验。我们测试了不同的SARS-CoV-2刺突蛋白肽、S1结构域和整个S1-S2胞外域，发现它们都没有以特异性方式明显胜过[125 I]-α-BgTx。此外，膜片钳记录显示S1结构域对α7-AChR介导的电流没有明显影响。我们的结论是，SARS-CoV-2刺突蛋白与人α7-AChR的正构位点的结合，以及它与乙酰胆碱、胆碱或尼古丁的竞争，不太可能是这种复杂疾病的相关方面。
Although much has been learned about the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the vast array of hitherto unexplained symptoms that characterize acute and postacute (“long”) COVID-19 warrants the search for additional biochemical pathways involved. On the basis of amino-acid sequence analyses and computational approaches, it has recently been suggested that nicotinic acetylcholine receptors (AChRs) may act as additional SARS-CoV-2 plasma membrane receptors through the binding of the virus’ spike protein to a site that overlaps with the orthosteric, ACh-binding sites. Here, on the basis of experimental ligand-binding competition assays, we conclude that the mutually exclusive binding of SARS-CoV-2 and cholinergic ligands to the human α7-AChR is unlikely to be a relevant aspect of this complex disease. The pathophysiological mechanisms underlying the constellation of symptoms that characterize COVID-19 are only incompletely understood. In an effort to fill these gaps, a “nicotinic hypothesis,” which posits that nicotinic acetylcholine receptors (AChRs) act as additional severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptors, has recently been put forth. A key feature of the proposal (with potential clinical ramifications) is the suggested competition between the virus’ spike protein and small-molecule cholinergic ligands for the receptor’s orthosteric binding sites. This notion is reminiscent of the well-established role of the muscle AChR during rabies virus infection. To address this hypothesis directly, we performed equilibrium-type ligand-binding competition assays using the homomeric human α7-AChR (expressed on intact cells) as the receptor, and radio-labeled α-bungarotoxin (α-BgTx) as the orthosteric-site competing ligand. We tested different SARS-CoV-2 spike protein peptides, the S1 domain, and the entire S1–S2 ectodomain, and found that none of them appreciably outcompete [125I]-α-BgTx in a specific manner. Furthermore, patch-clamp recordings showed no clear effect of the S1 domain on α7-AChR–mediated currents. We conclude that the binding of the SARS-CoV-2 spike protein to the human α7-AChR’s orthosteric sites—and thus, its competition with ACh, choline, or nicotine—is unlikely to be a relevant aspect of this complex disease.
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DOI：
10.1126/science.abd3072
发表时间：
2020-11-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Daly JL;Simonetti B;Klein K;Chen KE;Williamson MK;Antón-Plágaro C;Shoemark DK;Simón-Gracia L;Bauer M;Hollandi R;Greber UF;Horvath P;Sessions RB;Helenius A;Hiscox JA;Teesalu T;Matthews DA;Davidson AD;Collins BM;Cullen PJ;Yamauchi Y
通讯作者：
Yamauchi Y
SARS-COV-2尖峰蛋白的Y674-R685区与人α7烟碱受体之间的功能相互作用。
影响因子：
5.1
作者：
Facundo Chrestia, Juan;Oliveira, Ana Sofia;Mulholland, Adrian J.;Gallagher, Timothy;Bermudez, Isabel;Bouzat, Cecilia
通讯作者：
Bouzat, Cecilia
影响因子：
1.7
作者：
Bertrand, S;DevillersThiery, A;Bertrand, D
通讯作者：
Bertrand, D
影响因子：
3.6
作者：
Gronlien, Jens Halvard;Hakerud, Monika;Malysz, John
通讯作者：
Malysz, John
影响因子：
4.6
作者：
Alberto Verdin-Betancourt, Francisco;Figueroa, Mario;Sierra-Santoyo, Adolfo
通讯作者：
Sierra-Santoyo, Adolfo