Sagara J, Tochikura T S, Tanaka H, Baba Y, Tsukita S, Tsukita S, Kawai A
Department of Molecular Microbiology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Microbiol Immunol. 1998;42(4):289-97. doi: 10.1111/j.1348-0421.1998.tb02285.x.
In our monoclonal antibody (MAb) stocks prepared against the BHK-21 cell antigens, two (#11875 and 28276) recognized a 21-kDa polypeptide (referred to as VAP21) which is efficiently incorporated into the rabies virion. By using these MAbs, we isolated the cDNA clones that encoded a polypeptide of 144 amino acids from our BHK-21 cell cDNA library. Based on the following evidence, the cDNA was assumed to encode a full-length sequence of VAP21 antigen: i) expression of the cDNA in animal cells resulted in the production of a polypeptide recognized by the two MAbs, and its electrophoretic mobility was the same as that of authentic VAP21 antigen; and ii) immunization with the products from the cDNA-transformed E. coli cells raised specific antibodies in rabbits that recognized a 21-kDa polypeptide in the virion. From the deduced amino acid sequence, it is suggested that the VAP21 antigen has a molecular structure of type-I transmembrane protein containing characteristic proline-rich and glycine-rich regions in its ectodomain. Homology searches resulted in finding homologous sequences (totally about 40% homology) in the human MIC2 gene product (CD99; 32-kDa) of T lymphocytes. These results suggest that the VAP21 antigen in the rabies virion is a cellular CD99-related transmembrane protein.
在我们针对BHK - 21细胞抗原制备的单克隆抗体(MAb)储备中,两种抗体(#11875和28276)识别出一种21 kDa的多肽(称为VAP21),该多肽能有效掺入狂犬病毒粒子中。利用这些单克隆抗体,我们从BHK - 21细胞cDNA文库中分离出编码144个氨基酸多肽的cDNA克隆。基于以下证据,假定该cDNA编码VAP21抗原的全长序列:i)该cDNA在动物细胞中的表达产生了一种能被这两种单克隆抗体识别的多肽,其电泳迁移率与天然VAP21抗原相同;ii)用cDNA转化的大肠杆菌细胞产物免疫兔子,产生了能识别病毒粒子中21 kDa多肽的特异性抗体。从推导的氨基酸序列来看,VAP21抗原具有I型跨膜蛋白的分子结构,其胞外结构域含有特征性的富含脯氨酸和富含甘氨酸的区域。同源性搜索结果显示,在T淋巴细胞的人MIC2基因产物(CD99;32 kDa)中发现了同源序列(总体同源性约为40%)。这些结果表明,狂犬病毒粒子中的VAP21抗原是一种细胞CD99相关的跨膜蛋白。