Identification of a new region of SARS-CoV S protein critical for viral entry.

Identification of a new region of SARS-CoV S protein critical for viral entry.
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DOI:
10.1016/j.jmb.2009.10.032
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发表时间:
2009-12-11
影响因子:
5.6
通讯作者:
Caffrey M
Caffrey M
中科院分区:
生物学2区
文献类型:
--
作者:
Guo Y;Tisoncik J;McReynolds S;Farzan M;Prabhakar BS;Gallagher T;Rong L;Caffrey M

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严重急性呼吸综合征冠状病毒(SARS-CoV)的感染是通过SARS-CoV刺突蛋白(S)及其受体ACE2之间的特异性相互作用而启动的。在这份报告中,我们使用基于人类免疫缺陷病毒的伪分型系统筛选了一个代表SARS-CoV S蛋白序列的肽库,以确定影响病毒进入的特定区域。在筛选的169个肽中,9626肽(S残基217-234)以剂量依赖的方式抑制SARS-CoV S介导的假型病毒粒子进入表达功能性SARS-CoV受体(人血管紧张素转换酶2)的293T细胞(IC_(50)、∼和11、μ_M)。丙氨酸扫描突变被用来评估单个残基在S的这个区域内的作用,这是以前没有特征的。这些影响包括显著降低表达(K223A)、病毒掺入(L218A、I230A和N232A)和减少病毒进入(L224A、L226A、I228A、T231A和F233A)。综上所述,这些结果揭示了S蛋白的一个新区域,该区域对SARS-CoV的进入至关重要。
Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) is initiated by specific interactions between the SARS-CoV spike (S) protein and its receptor ACE2. In this report, we screened a peptide library representing the SARS-CoV S protein sequence using a human immunodeficiency virus-based pseudotyping system to identify specific regions that affect viral entry. One of the 169 peptides screened, peptide 9626 (S residues 217–234), inhibited SARS-CoV S-mediated entry of the pseudotyped virions in 293T cells expressing a functional SARS-CoV receptor (human angiotensin-converting enzyme 2) in a dose-dependent manner (IC50 ∼ 11 μM). Alanine scanning mutagenesis was performed to assess the roles of individual residues within this region of S, which was previously uncharacterized. The effects included significant reductions in expression (K223A), viral incorporation (L218A, I230A, and N232A), and reduced viral entry (L224A, L226A, I228A, T231A, and F233A). Taken together, these results reveal a new region of the S protein that is crucial for SARS-CoV entry.
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