Two Binding Sites of SARS-CoV-2 Macrodomain 3 Probed by Oxaprozin and Meclomen
Two Binding Sites of SARS-CoV-2 Macrodomain 3 Probed by Oxaprozin and Meclomen
复制标题
Oxaprozin 和 Meclomen 探测 SARS-CoV-2 宏结构域 3 的两个结合位点
DOI：
10.1021/acs.jmedchem.2c01168
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发表时间：
2022-11-10
影响因子：
7.3
通讯作者：
Gao，Jia
中科院分区：
文献类型：
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作者：
Li，Jiao;Zhong，Fumei;Gao，Jia
严重急性呼吸综合征冠状病毒1/2（SARS-CoV-1/2）宏结构域3（Mac 3）对病毒基因组的复制和转录至关重要，因此是潜在的治疗靶点。在这里，我们解决了SARS-CoV-2 Mac 3的晶体结构，揭示了一个小分子结合口袋。两种低分子量药物，奥沙普秦和美克氯明，诱导不同的模式的核磁共振（NMR）化学位移扰动（CSP）。Meclomen与SARS-CoV-2 Mac 3的位点I结合，结合位姿由NMR CSP和转移的顺磁弛豫增强确定，而奥沙普秦结合位点II，如晶体结构所示。有趣的是，奥沙普秦和meclomen都扰乱了SARS-CoV Mac 3的I位点残基。荧光偏振实验进一步证明奥沙普秦和美克洛门抑制DNA-G4 s与SARS-CoV-2 Mac 3的结合。我们的工作确定了SARS-CoV-2 Mac 3的两个相邻的配体结合位点，这将有助于这些化合物的结构指导的片段连接，以获得更有效的抑制剂。
Severe acute respiratory syndrome-coronavirus-1/2 (SARS-CoV-1/2) macrodomain 3 (Mac3) is critical for replication and transcription of the viral genome and is therefore a potential therapeutic target. Here, we solved the crystal structure of SARS-CoV-2 Mac3, which reveals a small-molecule binding pocket. Two low-molecular-weight drugs, oxaprozin and meclomen, induced different patterns of nuclear magnetic resonance (NMR) chemical shift perturbations (CSPs). Meclomen binds to site I of SARS-CoV-2 Mac3 with binding pose determined by NMR CSP and transferred paramagnetic relaxation enhancement, while oxaprozin binds to site II as revealed by the crystal structure. Interestingly, oxaprozin and meclomen both perturb residues in site I of SARS-CoV Mac3. Fluorescence polarization experiments further demonstrated that oxaprozin and meclomen inhibited the binding of DNA-G4s to SARS-CoV-2 Mac3. Our work identified two adjacent ligand-binding sites of SARS-CoV-2 Mac3 that shall facilitate structure-guided fragment linking of these compounds for more potent inhibitors.