Expression cloning of functional receptor used by SARS coronavirus.
Expression cloning of functional receptor used by SARS coronavirus.
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DOI：
10.1016/j.bbrc.2004.01.076
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发表时间：
2004-03-05
影响因子：
3.1
通讯作者：
Deng H
中科院分区：
文献类型：
--
作者：
Wang P;Chen J;Zheng A;Nie Y;Shi X;Wang W;Wang G;Luo M;Liu H;Tan L;Song X;Wang Z;Yin X;Qu X;Wang X;Qing T;Ding M;Deng H
我们表达了一系列 SARS-CoV 的截短刺突 (S) 糖蛋白，发现 N 末端 14-502 残基足以与 SARS-CoV 易感的 Vero E6 细胞结合。以该可溶性S蛋白片段作为亲和配体，筛选转染Vero E6细胞逆转录病毒cDNA文库的HeLa细胞，获得可与S蛋白结合的HeLa细胞克隆。该细胞克隆易受 HIV/SARS 假病毒感染，并且通过细胞-细胞融合测定证实该细胞克隆中存在 S 蛋白功能性受体。进一步的研究表明，该细胞的易感性是由于内源性血管紧张素转换酶 2 (ACE2) 的表达所致，该酶被用于表达克隆的逆转录病毒载体插入的 LTR 激活。当人ACE2 cDNA转导到NIH3T3细胞中时，表达ACE2的NIH3T3细胞可能会被HIV/SARS假病毒感染。这些数据清楚地表明 ACE2 是 SARS-CoV 的功能受体。
We have expressed a series of truncated spike (S) glycoproteins of SARS-CoV and found that the N-terminus 14–502 residuals were sufficient to bind to SARS-CoV susceptible Vero E6 cells. With this soluble S protein fragment as an affinity ligand, we screened HeLa cells transduced with retroviral cDNA library from Vero E6 cells and obtained a HeLa cell clone which could bind with the S protein. This cell clone was susceptible to HIV/SARS pseudovirus infection and the presence of a functional receptor for S protein in this cell clone was confirmed by the cell–cell fusion assay. Further studies showed the susceptibility of this cell was due to the expression of endogenous angiotensin-converting enzyme 2 (ACE2) which was activated by inserted LTR from retroviral vector used for expression cloning. When human ACE2 cDNA was transduced into NIH3T3 cells, the ACE2 expressing NIH3T3 cells could be infected with HIV/SARS pseudovirus. These data clearly demonstrated that ACE2 was the functional receptor for SARS-CoV.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
56.9
作者：
Pileri, P;Uematsu, Y;Abrignani, S
通讯作者：
Abrignani, S
DOI：
10.1016/j.bbrc.2003.11.054
发表时间：
2003-12-26
影响因子：
3.1
作者：
Xiao X;Chakraborti S;Dimitrov AS;Gramatikoff K;Dimitrov DS
通讯作者：
Dimitrov DS
影响因子：
7.2
作者：
Ponce, MR;Quesada, V;Micol, JL
通讯作者：
Micol, JL
影响因子：
64.5
作者：
Chan, SY;Empig, CJ;Goldsmith, MA
通讯作者：
Goldsmith, MA