Folding of the rabbit hemorrhagic disease virus capsid protein and delineation of N-terminal domains dispensable for assembly.
Folding of the rabbit hemorrhagic disease virus capsid protein and delineation of N-terminal domains dispensable for assembly.
复制标题
DOI：
10.1007/s00705-002-0825-3
复制
发表时间：
2002-08
影响因子：
2.7
通讯作者：
Rasschaert D
中科院分区：
文献类型：
--
作者：
Laurent S;Kut E;Remy-Delaunay S;Rasschaert D
兔出血性疾病病毒（RHDV）和欧洲褐兔综合症病毒（EBHSV）是杯状病毒，会产生严重的症状，对家兔和野兔具有致命性。通过确定由 RHDV 和 EBHSV 衣壳蛋白区域组成的嵌合衣壳蛋白的抗原模式来研究衣壳蛋白的折叠。抗 RHDV 单克隆抗体 (MAb) E3 已知可结合外部构象表位，可识别 RHDV C 末端区域。抗 RHDV MAb A47 结合埋藏表位，识别 RHDV N 末端区域。使用 pGEX 表达文库，我们更精确地将 MAb A47 表位定位在 31 个残基长度的肽上，位于 VP60 的残基 129 和 160 之间，确认其在蛋白质 N 端部分的位置。这些结果证明蛋白质的C端部分可与外部接触，而蛋白质的N端结构域构成颗粒的内壳结构域。为了利用 RHDV 的病毒样颗粒（VLP）作为表位载体或 DNA 转移载体，我们在杆状病毒系统中产生了三种 N 末端截短的蛋白质：ΔN1、ΔN2 和 ΔN3。 ΔN1 蛋白组装成 VLP，证明前 42 个氨基酸残基对于衣壳组装不是必需的。相比之下，缺失前 75 个残基的 ΔN2 无法形成 VLP。使用缺少残基 31 至 93（位于 RHDV 衣壳蛋白的免疫显性区域）的 ΔN3 蛋白获得的小颗粒表明，可以消除多达 62 个氨基酸残基而不妨碍组装。
Rabbit hemorrhagic disease virus (RHDV) and European brown hare syndrome virus (EBHSV) are caliciviruses that produce severe symptoms and are lethal to rabbits and hares. The folding of the capsid protein was studied by determination of the antigenic pattern of chimeric capsid proteins, composed of regions from RHDV and EBHSV capsid proteins. The anti-RHDV monoclonal antibody (MAb) E3, which is known to bind an external conformational epitope, recognized the RHDV C-terminal region. The anti-RHDV MAb A47, which binds a buried epitope, recognized the RHDV N-terminal region. Using a pGEX expression library, we more precisely mapped the MAb A47 epitope on a 31 residues length peptide, between residue 129 and 160 of the VP60, confirming its location in the N-terminal part of the protein. These results demonstrate that the C-terminal part of the protein is accessible to the exterior whereas the N-terminal domain of the protein constitutes the internal shell domain of the particle. With the aim of using virus-like particles (VLPs) of RHDV as epitope carriers or DNA transfer vectors, we produced in the baculovirus system three proteins, ΔN1, ΔN2 and ΔN3, truncated at the N terminus. The ΔN1 protein assembled into VLPs, demonstrating that the first 42 amino acid residues are not essential for capsid assembly. In contrast, ΔN2, from which the first 75 residues were missing, was unable to form VLPs. The small particles obtained with the ΔN3 protein lacking residues 31 to 93, located in the immunodominant region of the RHDV capsid protein, indicate that up to 62 amino acid residues can be eliminated without preventing assembly.
登录
查看更多内容
影响因子：
5
作者：
Capucci, L;Fallacara, F;Brocchi, E
通讯作者：
Brocchi, E
影响因子：
5.6
作者：
PRASAD, BVV;MATSON, DO;SMITH, AW
通讯作者：
SMITH, AW
影响因子：
5
作者：
Granzow, H;Weiland, F;Schirrmeier, H
通讯作者：
Schirrmeier, H
影响因子：
56.9
作者：
Prasad, BVV;Hardy, ME;Estes, MK
通讯作者：
Estes, MK
影响因子：
3.8
作者：
LeGall, G;Huguet, S;Rasschaert, D
通讯作者：
Rasschaert, D