Reconstitution of the receptor-binding motif of the SARS coronavirus
Reconstitution of the receptor-binding motif of the SARS coronavirus
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DOI：
10.1093/protein/gzv052
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发表时间：
2015-12-01
影响因子：
2.4
通讯作者：
Gershoni, Jonathan M.
中科院分区：
文献类型：
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作者：
Freund, Natalia T.;Roitburd-Berman, Anna;Gershoni, Jonathan M.
2003 年发现的严重急性呼吸综合征 (SARS) 冠状病毒 (CoV) 已感染全球约 8000 人，其中近 10% 死亡。 SARS 冠状病毒对靶细胞的感染是通过病毒刺突 (S) 蛋白（1255 个氨基酸）与其细胞受体血管紧张素转换酶 2 (ACE2) 的相互作用介导的。 SARS CoV 受体结合结构域（S 蛋白的氨基酸 N318-T509）沿其外围有一个延长的行程，与 ACE2 接触，并被指定为受体结合基序（RBM，氨基酸 S432-T486）。此外，RBM 是主要的抗原决定簇，能够引发中和抗体的产生。因此，RBM 的作用是一种双功能生物活性表面，可以通过抗体来证明，例如中和性人抗 SARS 单克隆抗体 (mAb) 80R，其靶向 RBM 并与 ACE2 受体竞争结合。在这里，我们利用噬菌体展示肽库来重建功能性 RBM。这是通过生成大量具有多种构象的候选 RBM 肽来实现的。用相应的配体筛选此类“Conformer Libraries”已产生短 RBM 构建体（约 40 个氨基酸），它可以结合 ACE2 受体和中和 mAb 80R。
The severe acute respiratory syndrome (SARS) coronavirus (CoV) identified in 2003 has infected similar to 8000 people worldwide, killing nearly 10% of them. The infection of target cells by the SARS CoV is mediated through the interaction of the viral Spike (S) protein (1255 amino acids) and its cellular receptor, angiotensin-converting enzyme 2 (ACE2). The SARS CoV receptor-binding domain (amino acids N318-T509 of S protein) harbors an extended excursion along its periphery that contacts ACE2 and is designated the receptor-binding motif (RBM, amino acids S432-T486). In addition, the RBM is a major antigenic determinant, able to elicit production of neutralizing antibodies. Hence, the role of the RBM is a bi-functional bioactive surface that can be demonstrated by antibodies such as the neutralizing human anti-SARS monoclonal antibody (mAb) 80R which targets the RBM and competes with the ACE2 receptor for binding. Here, we employ phage-display peptide-libraries to reconstitute a functional RBM. This is achieved by generating a vast collection of candidate RBM peptides that present a diversity of conformations. Screening such 'Conformer Libraries' with corresponding ligands has produced short RBM constructs (ca. 40 amino acids) that can bind both the ACE2 receptor and the neutralizing mAb 80R.