Antigenic and cellular localisation analysis of the severe acute respiratory syndrome coronavirus nucleocapsid protein using monoclonal antibodies.
Antigenic and cellular localisation analysis of the severe acute respiratory syndrome coronavirus nucleocapsid protein using monoclonal antibodies.
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DOI：
10.1016/j.virusres.2006.07.005
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发表时间：
2006-12
期刊：
影响因子：
5
通讯作者：
Jassoy C
中科院分区：
文献类型：
--
作者：
Bussmann BM;Reiche S;Jacob LH;Braun JM;Jassoy C
冠状病毒家族的一种成员此前已被确定为导致严重急性呼吸系统综合征（SARS）的原因。在这项研究中，已经产生了几种针对核衣壳蛋白的单克隆抗体，以检测核衣壳蛋白在病毒感染细胞中的分布，并研究该蛋白的抗原区域。共聚焦显微镜分析鉴定出包裹在核周区域囊泡中的核衣壳，表明病毒在内质网和高尔基体合成。单克隆抗体结合到核衣壳蛋白的中央和羧基端，表明该蛋白的这一部分具有突出的暴露性和免疫原性。抗体同时识别线性和构象表位。利用基于SARS核蛋白疏水性分析的数学模型预测抗原性不能得到充分证实。与不连续肽结合的抗体证明，氨基酸274-283和373-382组装成一个特别富含碱性氨基酸的结构单元。此外，代表单克隆抗体6D11C1识别的表位的286-295、316-325和361-367氨基酸会聚在一起，表明SARS病毒核衣壳蛋白的c端结构良好，以及所涉及的肽区域的功能关系。或者，核衣壳蛋白的二聚化可导致一个核蛋白分子上的氨基酸序列316-325和361-367与第二个肽上的氨基酸286-295并置。该单克隆抗体将可用于评估不同SARS冠状病毒株之间的抗原性和免疫学变异。
A member of the family of coronaviruses has previously been identified as the cause of the severe acute respiratory syndrome (SARS). In this study, several monoclonal antibodies against the nucleocapsid protein have been generated to examine distribution of the nucleocapsid in virus-infected cells and to study antigenic regions of the protein. Confocal microscopic analysis identified nucleocapsids packaged in vesicles in the perinuclear area indicating viral synthesis at the endoplasmic reticulum and Golgi apparatus. The monoclonal antibodies bound to the central and carboxyterminal half of the nucleocapsid protein indicating prominent exposure and immunogenicity of this part of the protein. Antibodies recognised both linear and conformational epitopes. Predictions of antigenicity using mathematical modelling based on hydrophobicity analysis of SARS nucleoprotein could not be confirmed fully. Antibody binding to discontinuous peptides provides evidence that amino acids 274–283 and 373–382 assemble to a structural unit particularly rich in basic amino acids. In addition, amino acids 286–295, 316–325 and 361–367 that represent the epitope recognised by monoclonal antibody 6D11C1 converge indicating a well-structured C-terminal region of the SARS virus nucleocapsid protein and functional relationship of the peptide regions involved. Alternatively, dimerisation of the nucleocapsid protein may result in juxtaposition of the amino acid sequences 316–325 and 361–367 on one nucleoprotein molecule to amino acid 286–295 on the second peptide. The monoclonal antibodies will be available to assess antigenicity and immunological variabilities between different SARS CoV strains.
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DOI：
10.1016/j.str.2005.08.021
发表时间：
2005-12
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Fan H;Ooi A;Tan YW;Wang S;Fang S;Liu DX;Lescar J
通讯作者：
Lescar J
影响因子：
3.8
作者：
Nelson, GW;Stohlman, SA;Tahara, SM
通讯作者：
Tahara, SM
影响因子：
2.9
作者：
Huang, QL;Yu, LP;Olejniczak, ET
通讯作者：
Olejniczak, ET
影响因子：
5.4
作者：
Rowland, RRR;Chauhan, V;Burton, MD
通讯作者：
Burton, MD
影响因子：
5
作者：
Timani KA;Liao Q;Ye L;Zeng Y;Liu J;Zheng Y;Ye L;Yang X;Lingbao K;Gao J;Zhu Y
通讯作者：
Zhu Y