Hexamethylene amiloride binds the SARS-CoV-2 envelope protein at the protein-lipid interface.
Hexamethylene amiloride binds the SARS-CoV-2 envelope protein at the protein-lipid interface.
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DOI：
10.1002/pro.4755
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发表时间：
2023-10
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影响因子：
--
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中科院分区：
文献类型：
--
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SARS-COV-2蛋白质在脂质双层中形成五螺旋束，其阳离子传导活性与炎症反应和呼吸道遇险症状有关。 HMA和E跨膜之间脂质双层中的域（ETM）与15n标记的HMA与氟化或13C标记的ETM结合使用。 MA-蛋白距离。每个五聚会的药物都意外地位于通道孔内，但在TM域中的脂质面孔表面上，HMA可以通过在较低的毒品下persitions persitions persitions persitions，这表明HMA可能会抑制E通道的活性。这种差异表明，与通道孔相比，HMA对蛋白质 - 脂质界面具有更高的亲和力。
The SARS‐CoV‐2 envelope (E) protein forms a five‐helix bundle in lipid bilayers whose cation‐conducting activity is associated with the inflammatory response and respiratory distress symptoms of COVID‐19. E channel activity is inhibited by the drug 5‐(N,N‐hexamethylene) amiloride (HMA). However, the binding site of HMA in E has not been determined. Here we use solid‐state NMR to measure distances between HMA and the E transmembrane domain (ETM) in lipid bilayers. 13C, 15N‐labeled HMA is combined with fluorinated or 13C‐labeled ETM. Conversely, fluorinated HMA is combined with 13C, 15N‐labeled ETM. These orthogonal isotopic labeling patterns allow us to conduct dipolar recoupling NMR experiments to determine the HMA binding stoichiometry to ETM as well as HMA‐protein distances. We find that HMA binds ETM with a stoichiometry of one drug per pentamer. Unexpectedly, the bound HMA is not centrally located within the channel pore, but lies on the lipid‐facing surface in the middle of the TM domain. This result suggests that HMA may inhibit the E channel activity by interfering with the gating function of an aromatic network. These distance data are obtained under much lower drug concentrations than in previous chemical shift perturbation data, which showed the largest perturbation for N‐terminal residues. This difference suggests that HMA has higher affinity for the protein–lipid interface than the channel pore. These results give insight into the inhibition mechanism of HMA for SARS‐CoV‐2 E.
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影响因子：
2.2
作者：
Hou, Guangjin;Yan, Si;Trebosc, Julien;Amoureux, Jean-Paul;Polenova, Tatyana
通讯作者：
Polenova, Tatyana
影响因子：
7.3
作者：
Buckley BJ;Aboelela A;Minaei E;Jiang LX;Xu Z;Ali U;Fildes K;Cheung CY;Cook SM;Johnson DC;Bachovchin DA;Cook GM;Apte M;Huang M;Ranson M;Kelso MJ
通讯作者：
Kelso MJ
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
4.9
作者：
Ewart, GD;Nasr, N;Gage, PW
通讯作者：
Gage, PW
影响因子：
2.9
作者：
Andreas, Loren B.;Barnes, Alexander B.;Griffin, Robert G.
通讯作者：
Griffin, Robert G.