1H, 13C and 15N resonance assignment of the SARS-CoV-2 full-length nsp1 protein and its mutants reveals its unique secondary structure features in solution.
1H, 13C and 15N resonance assignment of the SARS-CoV-2 full-length nsp1 protein and its mutants reveals its unique secondary structure features in solution.
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DOI：
10.1371/journal.pone.0251834
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Agback P
中科院分区：
文献类型：
--
作者：
Agback T;Dominguez F;Frolov I;Frolova EI;Agback P
SARS-CoV-2全长nsp1蛋白的结构表征将成为开发针对SARS-CoV-2的靶向抗病毒药物的重要工具，并有助于进一步了解该蛋白的分子内和分子间相互作用。作为该蛋白核磁共振研究的第一步，我们报告了全长sars - cov -2 nsp1的1H， 13C和15N共振主链分配以及载子形式的Cβ，包括折叠结构域以及片状的N-和C-末端内在无序片段。该蛋白为19.8 kD，经高分辨率NMR表征。利用两个不同的突变体H81P和K129E/D48E以及氨基酸特异性实验验证了分配的有效性。根据得到的赋值，溶液中折叠畴的二级结构与先前发表的x射线结构以及另一种已发表的NMR得到的二级结构几乎相同，但发现了一些差异。在溶液中，SARS-CoV-2 nsp1表现出无序、柔性的N端和c端，具有不同的动态特性。无序c端结构域起始的短肽在NMR时间尺度上可区分为两种不同的构象。我们认为无序和折叠的nsp1结构域不是完全独立的单元，而是参与分子内相互作用。对溶液中SARS-CoV-2突变体的结构和动力学的研究正在进行中，这将为这些相互作用背后的分子机制提供重要见解。
Structural characterization of the SARS-CoV-2 full length nsp1 protein will be an essential tool for developing new target-directed antiviral drugs against SARS-CoV-2 and for further understanding of intra- and intermolecular interactions of this protein. As a first step in the NMR studies of the protein, we report the 1H, 13C and 15N resonance backbone assignment as well as the Cβ of the apo form of the full-lengthSARS-CoV-2 nsp1 including the folded domain together with the flaking N- and C- terminal intrinsically disordered fragments. The 19.8 kD protein was characterized by high-resolution NMR. Validation of assignment have been done by using two different mutants, H81P and K129E/D48E as well as by amino acid specific experiments. According to the obtained assignment, the secondary structure of the folded domain in solution was almost identical to its previously published X-ray structure as well as another published secondary structure obtained by NMR, but some discrepancies have been detected. In the solution SARS-CoV-2 nsp1 exhibited disordered, flexible N- and C-termini with different dynamic characteristics. The short peptide in the beginning of the disordered C-terminal domain adopted two different conformations distinguishable on the NMR time scale. We propose that the disordered and folded nsp1 domains are not fully independent units but are rather involved in intramolecular interactions. Studies of the structure and dynamics of the SARS-CoV-2 mutant in solution are on-going and will provide important insights into the molecular mechanisms underlying these interactions.
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影响因子：
5.8
作者：
Semper C;Watanabe N;Savchenko A
通讯作者：
Savchenko A
影响因子：
6.7
作者：
Huang C;Lokugamage KG;Rozovics JM;Narayanan K;Semler BL;Makino S
通讯作者：
Makino S
影响因子：
3.7
作者：
Brockway SM;Denison MR
通讯作者：
Denison MR
DOI：
10.1073/pnas.0603144103
发表时间：
2006-08-22
影响因子：
11.1
作者：
Kamitani, Wataru;Narayanan, Krishna;Makino, Shinji
通讯作者：
Makino, Shinji
DOI：
10.1046/j.1432-1327.1998.2560001.x
发表时间：
1998-08-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Markley, JL;Bax, A;Wüthrich, K
通讯作者：
Wüthrich, K