Tomassini J E, Graham D, DeWitt C M, Lineberger D W, Rodkey J A, Colonno R J
Department of Virus and Cell Biology, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Proc Natl Acad Sci U S A. 1989 Jul;86(13):4907-11. doi: 10.1073/pnas.86.13.4907.
A 90-kDa surface glycoprotein was previously isolated and shown to be required for infection by the "major" group of human rhinovirus (HRV) serotypes. In the present work, the amino acid sequence of the receptor protein was obtained from CNBr and tryptic peptides. Using degenerate oligonucleotides predicted from the peptide sequences, we identified four cDNA clones that encode a 3-kilobase mRNA. The clones were ligated, subcloned in a simian virus 40 expression vector, and used to transfect receptor-negative Vero (monkey) cells. Results showed that transfected cells expressed receptor molecules capable of binding HRV and a monoclonal antibody which recognizes the major group HRV receptor. The cloned receptor cDNA encoded a protein with a sequence nearly identical to that of the intercellular adhesion molecule 1 (ICAM-1), indicating that the two surface proteins are one and the same. Both proteins have identical mass, carbohydrate composition, and tissue distribution. In addition, major group receptors on HeLa cells could be induced with various cytokines in a manner similar to the ICAM-1 ligand. A similar induction of the HRV "minor" group receptor was not observed.
一种90kDa的表面糖蛋白先前已被分离出来，并被证明是人类鼻病毒（HRV）“主要”血清型感染所必需的。在本研究中，通过溴化氰和胰蛋白酶肽段获得了受体蛋白的氨基酸序列。利用从肽序列预测的简并寡核苷酸，我们鉴定出四个编码3千碱基mRNA的cDNA克隆。将这些克隆连接起来，亚克隆到猿猴病毒40表达载体中，并用于转染缺乏受体的非洲绿猴肾（Vero）细胞。结果表明，转染后的细胞表达出能够结合HRV的受体分子以及一种识别主要组HRV受体的单克隆抗体。克隆的受体cDNA编码的蛋白质序列与细胞间黏附分子1（ICAM-1）的序列几乎相同，表明这两种表面蛋白是同一物质。这两种蛋白具有相同的分子量、碳水化合物组成和组织分布。此外，HeLa细胞上的主要组受体可以被多种细胞因子以类似于ICAM-1配体的方式诱导。未观察到HRV“次要”组受体的类似诱导现象。
