Receptor-binding ability of fragments 260-600 and 397-796 of SARS-associated coronavirus spike protein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 353-357, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-333007
ABSTRACT
<p><b>BACKGROUND</b>To investigate the interaction between the host cell and the truncated S fragments to identify the receptor-binding domain of the spike (S) protein of SARS-associated coronavirus (SARS-CoV).</p><p><b>METHODS</b>Two different fragments S260-600 and S397-796 of the SARS-CoV S protein were expressed in Escherichia coli (E.coli) using a pET expression vector, respectively. The two recombinant proteins were separately verified by Western blot, purified by nickel-affinity chromatography, and incubated with Vero cells, a susceptible cell line of SARS-CoV infection, for cell binding assay. After the sequential probing with sera from convalescent SARS-patients and FITC-labeled anti-human IgG, the cells were analyzed by flow cytometry. The NIH 3T3 cell, a non-permissive cell line of SARS-CoV infection, was used as controls.</p><p><b>RESULTS</b>The recombinant proteins S260-600 and S397-796 were efficiently expressed in an insoluble form in E.coli. The appropriate expression of the proteins was confirmed by Western blotting using both SARS patients' sera and anti-6 x histidine antibody. The flow cytometry results showed that the both proteins were able to bind Vero cells, but the binding ability of S260-600 was somewhat stronger than that of S397-796. In contrast, the S260-600 protein did not bind NIH3T3 cells.</p><p><b>CONCLUSION</b>Both S260-600 and S397-796 exhibited different receptor binding activity. The S260-600 fragment probably contains the important receptor binding domain and could be a potential candidate for the development of SARS vaccine and anti-SARS therapeutics.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Fragmentos de Peptídeos
/
Ligação Proteica
/
Ligação Competitiva
/
Células Vero
/
Proteínas Recombinantes
/
Glicoproteínas de Membrana
/
Chlorocebus aethiops
/
Química
/
Proteínas do Envelope Viral
/
Western Blotting
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2005
Tipo de documento:
Artigo
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