Identification of fragments binding to SARS-CoV-2 nsp10 reveals ligand-binding sites in conserved interfaces between nsp10 and nsp14/nsp16.
Identification of fragments binding to SARS-CoV-2 nsp10 reveals ligand-binding sites in conserved interfaces between nsp10 and nsp14/nsp16.
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DOI：
10.1039/d1cb00135c
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发表时间：
2022-01-05
影响因子：
4.1
通讯作者：
Knecht W
中科院分区：
文献类型：
--
作者：
Kozielski F;Sele C;Talibov VO;Lou J;Dong D;Wang Q;Shi X;Nyblom M;Rogstam A;Krojer T;Fisher Z;Knecht W
自 2019 年 SARS-CoV-2 出现以来，Covid-19 已发展成为对我们的健康、社会和经济系统的严重威胁。尽管疫苗的开发已取得了惊人的成果，并且现在越来越容易获得，但现有药物的重新利用却不太成功。显然需要开发针对 SARS-CoV-2 的新药物，这些药物也可用于对抗未来的冠状病毒感染。非结构蛋白 10 (nsp10) 是两种对病毒复制至关重要的酶 nsp14 和 nsp16 的保守刺激物，具有核糖核酸外切酶和甲基转移酶活性。干扰 RNA 校对或 RNA 帽形成代表了抑制复制的干预策略。我们使用纳米差示扫描荧光测定法和 X 射线晶体学进行基于片段的筛选，以识别针对 SARS-CoV-2 nsp10 的配体。我们确定了位于两个不同位点的四个片段：一个可以建模为它位于 nsp14-nsp10 复合物界面中的位置，另一个位于 nsp16-nsp10 复合物界面中。使用微尺度热泳 (MST) 实验来量化 nsp10 的片段亲和力。此外，我们通过 MST 表明 nsp14 和 10 的相互作用很弱，因此小分子可能会破坏复合物的形成。这些片段将作为使用片段生长技术和基于结构的药物设计开发更有效的类似物的起点。通过使用 X 射线晶体学的片段筛选，我们鉴定了四种配体，揭示了 SARS-CoV-2 nsp10 和 nsp14/nsp16 之间界面的配体结合位点。 nsp14/10 相互作用较弱，因此可能会被小分子破坏。
Since the emergence of SARS-CoV-2 in 2019, Covid-19 has developed into a serious threat to our health, social and economic systems. Although vaccines have been developed in a tour-de-force and are now increasingly available, repurposing of existing drugs has been less successful. There is a clear need to develop new drugs against SARS-CoV-2 that can also be used against future coronavirus infections. Non-structural protein 10 (nsp10) is a conserved stimulator of two enzymes crucial for viral replication, nsp14 and nsp16, exhibiting exoribonuclease and methyltransferase activities. Interfering with RNA proofreading or RNA cap formation represents intervention strategies to inhibit replication. We applied fragment-based screening using nano differential scanning fluorometry and X-ray crystallography to identify ligands targeting SARS-CoV-2 nsp10. We identified four fragments located in two distinct sites: one can be modelled to where it would be located in the nsp14–nsp10 complex interface and the other in the nsp16–nsp10 complex interface. Microscale thermophoresis (MST) experiments were used to quantify fragment affinities for nsp10. Additionally, we showed by MST that the interaction by nsp14 and 10 is weak and thereby that complex formation could be disrupted by small molecules. The fragments will serve as starting points for the development of more potent analogues using fragment growing techniques and structure-based drug design. By fragment screening using X-ray crystallography we identified four ligands revealing ligand-binding sites in interfaces between SARS-CoV-2 nsp10 and nsp14/nsp16. The nsp14/10 interaction is weak and therefore could be disrupted by small molecules.
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影响因子：
5.6
作者：
Rogstam A;Nyblom M;Christensen S;Sele C;Talibov VO;Lindvall T;Rasmussen AA;André I;Fisher Z;Knecht W;Kozielski F
通讯作者：
Kozielski F
DOI：
10.1107/s2059798321003818
发表时间：
2021-06-01
期刊：
Acta crystallographica. Section D, Structural biology
影响因子：
--
作者：
Lima GMA;Jagudin E;Talibov VO;Benz LS;Marullo C;Barthel T;Wollenhaupt J;Weiss MS;Mueller U
通讯作者：
Mueller U
DOI：
10.1107/s2059798317000067
发表时间：
2017-02-01
期刊：
Acta crystallographica. Section D, Structural biology
影响因子：
--
作者：
Long F;Nicholls RA;Emsley P;Graǽulis S;Merkys A;Vaitkus A;Murshudov GN
通讯作者：
Murshudov GN
影响因子：
--
作者：
Chatrin C;Talapatra SK;Canard B;Kozielski F
通讯作者：
Kozielski F
影响因子：
64.8
作者：
通讯作者：
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