The SARS-CoV S glycoprotein: expression and functional characterization.
The SARS-CoV S glycoprotein: expression and functional characterization.
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DOI：
10.1016/j.bbrc.2003.11.054
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发表时间：
2003-12-26
影响因子：
3.1
通讯作者：
Dimitrov DS
中科院分区：
文献类型：
--
作者：
Xiao X;Chakraborti S;Dimitrov AS;Gramatikoff K;Dimitrov DS
我们克隆、表达了SARS-CoV （Tor2分离物）S糖蛋白的全长和各种可溶性片段，并对其进行了鉴定。表达S的细胞在中性pH下与表达受体的细胞融合，这表明重组糖蛋白是功能性的，其膜融合活性不需要其他病毒蛋白，也不需要低pH来触发膜融合；在低受体浓度下未观察到融合。S及其可溶性外结构域Se没有明显的裂解。它们在SDS凝胶中约为180-200 kDa，这表明与之前的计算机分析预测的翻译后修饰和对其他冠状病毒的观察结果一致。含有n端氨基酸残基17-537和272-537的片段，但不含17-276的片段，与Vero E6细胞和纯化的可溶性受体ACE2特异性结合，最近被M. Farzan和同事发现[Nature 426(2003) 450-454]。结合针对S肽开发的抗体抑制结合的数据，这些发现表明受体结合域位于氨基酸残基303和537之间。这些结果也证实了ACE2是SARS病毒的功能性受体，可能有助于阐明SARS- cov进入机制以及疫苗免疫原和进入抑制剂的开发。
We have cloned, expressed, and characterized the full-length and various soluble fragments of the SARS-CoV (Tor2 isolate) S glycoprotein. Cells expressing S fused with receptor-expressing cells at neutral pH suggesting that the recombinant glycoprotein is functional, its membrane fusogenic activity does not require other viral proteins, and that low pH is not required for triggering membrane fusion; fusion was not observed at low receptor concentrations. S and its soluble ectodomain, Se, were not cleaved to any significant degree. They ran at about 180–200 kDa in SDS gels suggesting post-translational modifications as predicted by previous computer analysis and observed for other coronaviruses. Fragments containing the N-terminal amino acid residues 17–537 and 272–537 but not 17–276 bound specifically to Vero E6 cells and purified soluble receptor, ACE2, recently identified by M. Farzan and co-workers [Nature 426 (2003) 450–454]. Together with data for inhibition of binding by antibodies developed against peptides from S, these findings suggest that the receptor-binding domain is located between amino acid residues 303 and 537. These results also confirm that ACE2 is a functional receptor for the SARS virus and may help in the elucidation of the mechanisms of SARS-CoV entry and in the development of vaccine immunogens and entry inhibitors.
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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：
64.5
作者：
Dimitrov, DS
通讯作者：
Dimitrov, DS
影响因子：
3.7
作者：
Gallagher TM;Buchmeier MJ
通讯作者：
Buchmeier MJ
影响因子：
3.7
作者：
KUNKEL, F;HERRLER, G
通讯作者：
HERRLER, G
影响因子：
5.4
作者：
Bonavia, A;Zelus, BD;Holmes, KV
通讯作者：
Holmes, KV