Characterization of the interaction of domain III of the envelope protein of dengue virus with putative receptors from CHO cells
Characterization of the interaction of domain III of the envelope protein of dengue virus with putative receptors from CHO cells
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DOI：
10.1016/j.virusres.2008.07.022
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发表时间：
2008-11-01
期刊：
影响因子：
5
通讯作者：
Padron, Gabriel
中科院分区：
文献类型：
--
作者：
Huerta, Vivian;Chinea, Glay;Padron, Gabriel
登革热病毒 (DENV) 包膜蛋白的结构域 III (DIII) 包含与细胞受体相互作用的结构决定因素。在本研究中，使用固相测定和含有血清型 1 和 2 的 DENV-DIII 的重组融合蛋白来研究包膜蛋白与 CHO 细胞微粒体部分中存在的推定受体相互作用的结构特征。重组融合蛋白显示出与微粒体部分中存在的蛋白质的特异性相互作用。融合蛋白在 pH 5.5-8.0 范围内的结合类似于病毒颗粒，在 pH 6.0 时达到峰值。这表明莳萝与细胞受体的相互作用在内体 pH 值下得到加强。半胱氨酸残基的还原和脲甲基化对微粒体部分的结合和抗体识别的影响表明，与推定受体相互作用的 DIII 区域仅部分与抗体反应的主要表位重叠。残基保守谱的分析表明，莳萝的表面通常由特定的亚复合残基组成，在表面发现与主要中和表位密切相关的特定类型/亚型残基的代表性增加。 (c) 2008 Elsevier B.V. 保留所有权利。
Domain III (DIII) of the envelope protein of dengue virus (DENV) contains structural determinants for the interaction with cellular receptors. In the present study a solid phase assay and recombinant fusion proteins containing DENV-DIII of serotypes 1 and 2 were used to study structural features of the interaction of the envelope protein with putative receptors present in the microsomal fraction of CHO cells. Recombinant fusion proteins showed specific interaction with proteins present in the microsomal fraction. Binding of the fusion proteins across the pH range of 5.5-8.0 resembled that of virus particles, peaking at pH 6.0. This suggests that the interaction of Dill with cell receptor(s) is strengthened at endosomal pH. The effect of reduction and carbamidomethylation of cysteine residues on the binding to the microsomal fraction and in their recognition by antibodies suggests that the region of DIII that is interacting with putative receptor(s) overlaps only partially with a dominant epitope of the antibody response. The analysis of the residue conservation profile indicates that the surface of Dill is composed typically of specific subcomplex residues with an increased representation of specific type/subtype residues found at the surface that closely correlates with the dominant neutralizing epitope. (c) 2008 Elsevier B.V. All rights reserved.