Structure-based design of prefusion-stabilized SARS-CoV-2 spikes
Structure-based design of prefusion-stabilized SARS-CoV-2 spikes
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DOI：
10.1126/science.abd0826
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发表时间：
2020-09-18
期刊：
影响因子：
56.9
通讯作者：
McLellan, Jason S.
中科院分区：
文献类型：
--
作者：
Hsieh, Ching-Lin;Goldsmith, Jory A.;McLellan, Jason S.
2019年冠状病毒病（COVID-19）大流行导致了开发治疗剂和疫苗的加速努力。这些努力的一个关键目标是尖峰蛋白，它是可稳定且难以重组产生的。我们表征了100种结构引导的尖峰设计，并确定了26个单个取代，以增加蛋白质的产量和稳定性。有益替代的测试组合导致己糖的鉴定，己糖的识别是一种具有六个有益脯氨酸替代的变体，表现出更高的表达，比其父母构造（以10倍为10），并且能够承受热应力，在室温下存储和三个能力冻融周期。己糖在3.2的分辨率下的冷冻电子显微镜结构证实，它保留了预选尖峰构象。稳定的预融合峰值蛋白的高收益产生将加速疫苗和严重急性呼吸综合征冠状病毒2（SARS-COV-2）的疫苗和血清学诊断。
The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability. Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).