D614G Mutation Alters SARS-CoV-2 Spike Conformation and Enhances Protease Cleavage at the S1/S2 Junction.
D614G Mutation Alters SARS-CoV-2 Spike Conformation and Enhances Protease Cleavage at the S1/S2 Junction.
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DOI：
10.1016/j.celrep.2020.108630
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发表时间：
2021-01-12
期刊：
影响因子：
8.8
通讯作者：
Acharya P
中科院分区：
文献类型：
--
作者：
Gobeil SM;Janowska K;McDowell S;Mansouri K;Parks R;Manne K;Stalls V;Kopp MF;Henderson R;Edwards RJ;Haynes BF;Acharya P
严重急性呼吸道冠状病毒2型（SARS-CoV-2）刺突（S）蛋白是疫苗设计工作的目标，旨在结束2019年冠状病毒病（COVID-19）大流行。尽管突变率较低，但S蛋白中D 614 G取代的分离株在大流行期间出现较早，现在是全球的主要形式。在这里，我们探讨S构象的变化和D 614 G突变对可溶性S胞外域构建的影响。冷冻电子显微镜（cryo-EM）结构显示改变受体结合结构域（RBD）的处置;抗原性和蛋白水解实验揭示结构变化和增强弗林蛋白酶切割效率的G614变体。此外，弗林蛋白酶切割改变了G614 S胞外域中RBD的上/下比率，证明了对RBD定位的变构效应，该变构效应由SD 2区的变化触发，SD 2区含有残基614和弗林蛋白酶切割位点。我们的结果阐明了SARS-CoV-2 S的构象格局和变构性，并对疫苗设计具有影响。SARS-CoV-2 S 2 P突变不影响其结构、稳定性或抗原性D 614 G突变增加RBD“向上”状态并增强S1/S2连接蛋白水解结构和抗原性揭示S1/S2连接和RBD之间的变构SD 2锚定移动的RBD和NTD，将大的S1亚基运动与S2刺突分离SARS-CoV-2在细胞融合期间经历大的构象变化。Gobeil等人鉴定亚结构域锚，其限制融合前刺突的受体结合亚基中的大运动传播到其融合亚基。他们证明D 614 G突变增加了弗林蛋白酶切割的速率，这可能会影响感染性。
The severe acute respiratory coronavirus 2 (SARS-CoV-2) spike (S) protein is the target of vaccine design efforts to end the coronavirus disease 2019 (COVID-19) pandemic. Despite a low mutation rate, isolates with the D614G substitution in the S protein appeared early during the pandemic and are now the dominant form worldwide. Here, we explore S conformational changes and the effects of the D614G mutation on a soluble S ectodomain construct. Cryoelectron microscopy (cryo-EM) structures reveal altered receptor binding domain (RBD) disposition; antigenicity and proteolysis experiments reveal structural changes and enhanced furin cleavage efficiency of the G614 variant. Furthermore, furin cleavage alters the up/down ratio of the RBDs in the G614 S ectodomain, demonstrating an allosteric effect on RBD positioning triggered by changes in the SD2 region, which harbors residue 614 and the furin cleavage site. Our results elucidate SARS-CoV-2 S conformational landscape and allostery and have implications for vaccine design. SARS-CoV-2 S 2P mutations do not impact its structure, stability, or antigenicity D614G mutation increases RBD “up” state and enhances S1/S2 junction proteolysis Structure and antigenicity reveal allostery between the S1/S2 junction and RBD SD2 anchors the mobile RBD and NTD, separating large S1 subunit motions from S2 SARS-CoV-2 spike undergoes large conformational changes during cell fusion. Gobeil et al. identify a subdomain anchor that limits large motions in the receptor binding subunit of the pre-fusion spike from propagating to its fusion subunit. They demonstrate that the D614G mutation increases the rate of furin cleavage, which may impact infectivity.
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发表时间：
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期刊：
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