Dichlorophenylpyridine-Based Molecules Inhibit Furin through an Induced-Fit Mechanism.
Dichlorophenylpyridine-Based Molecules Inhibit Furin through an Induced-Fit Mechanism.
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DOI：
10.1021/acschembio.2c00103
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发表时间：
2022-04-15
影响因子：
4
通讯作者：
Brandstetter, Hans
中科院分区：
文献类型：
--
作者：
Dahms, Sven O.;Schnapp, Gisela;Winter, Martin;Buttner, Frank H.;Schlepuetz, Marco;Gnamm, Christian;Pautsch, Alexander;Brandstetter, Hans
前蛋白转化酶弗林蛋白酶的抑制剂可能作为广谱抗病毒治疗。已经报道了（3，5-二氯苯基）吡啶衍生的弗林蛋白酶抑制剂对急性呼吸综合征冠状病毒2（SARS-CoV-2）的高细胞效力和抗病毒活性。在这里，我们的特点是这种抑制剂类的结合机制，使用结构，生物物理和生物化学方法。我们建立了一个基于MALDI-TOF-MS的弗林蛋白酶活性测定，确定IC 50值，并解决了与弗林蛋白酶复合的（3，5-二氯苯基）吡啶衍生化合物的X-射线结构。抑制剂诱导的活性位点裂缝暴露一个中央掩埋色氨酸残基的大量构象重排。这些变化形成了一个扩展的疏水表面补丁的抑制剂的3，5-二氯苯基部分插入到一个新形成的结合口袋。与这些结构重排一致，我们分别在表面等离子体共振和差示扫描荧光实验中观察到缓慢的解离速率结合动力学和强的结构稳定性。发现的弗林蛋白酶构象为基于结构的药物发现提供了新的机会。
Inhibitors of the proprotein convertase furin might serve as broad-spectrum antiviral therapeutics. High cellular potency and antiviral activity against acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been reported for (3,5-dichlorophenyl)pyridine-derived furin inhibitors. Here we characterized the binding mechanism of this inhibitor class using structural, biophysical, and biochemical methods. We established a MALDI-TOF-MS-based furin activity assay, determined IC50 values, and solved X-ray structures of (3,5-dichlorophenyl)pyridine-derived compounds in complex with furin. The inhibitors induced a substantial conformational rearrangement of the active-site cleft by exposing a central buried tryptophan residue. These changes formed an extended hydrophobic surface patch where the 3,5-dichlorophenyl moiety of the inhibitors was inserted into a newly formed binding pocket. Consistent with these structural rearrangements, we observed slow off-rate binding kinetics and strong structural stabilization in surface plasmon resonance and differential scanning fluorimetry experiments, respectively. The discovered furin conformation offers new opportunities for structure-based drug discovery.
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影响因子：
2.7
作者：
Winter, Martin;Ries, Robert;Buettner, Frank H.
通讯作者：
Buettner, Frank H.
影响因子：
3.4
作者：
Hardes, Kornelia;Becker, Gero L.;Steinmetzer, Torsten
通讯作者：
Steinmetzer, Torsten
DOI：
10.1038/nsb941
发表时间：
2003-07-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Henrich, S;Cameron, A;Than, ME
通讯作者：
Than, ME
影响因子：
4.8
作者：
Cameron, A;Appel, J;Lindberg, I
通讯作者：
Lindberg, I
影响因子：
3.1
作者：
Winter, Martin;Bretschneider, Tom;Buettner, Frank H.
通讯作者：
Buettner, Frank H.