Association of ganglioside-protein conjugates into cell and Sendai virus. Requirement for the HN subunit in viral fusion.
Association of ganglioside-protein conjugates into cell and Sendai virus. Requirement for the HN subunit in viral fusion.
复制标题
神经节苷脂-蛋白质缀合物与细胞和仙台病毒的结合。
DOI：
10.1016/0014-4827(83)90389-0
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发表时间：
1983
影响因子：
3.7
通讯作者：
Macher,BA
中科院分区：
文献类型：
--
作者：
Heath,TD;Martin,FJ;Macher,BA
描述了一种制备蛋白质，特别是抗体及其片段与胶束形式的神经节苷脂的共价缀合物的方法。蛋白质-神经节苷脂缀合物与神经节苷脂胶束缔合，并且可以通过分子筛色谱与游离蛋白质分离。结合物可以不可逆地从胶束转移到所选择的细胞膜上，并且蛋白质部分被鉴定为新的表面抗原。这种方法的成功应用已被证明与三个生物系统。将兔IgG-神经节苷脂结合物转移到人或绵羊红细胞中，这些红细胞已与山羊抗兔IgG发生血凝。用神经节苷脂-抗H2KK修饰的红细胞已被证明粘附于表达H2KK抗原的L929小鼠成纤维细胞的单层。小鼠单克隆抗血型糖蛋白神经节苷脂结合物可与仙台病毒结合，并赋予病毒凝集和溶血去唾液酸化人红细胞的能力。使用抗血型糖蛋白缀合物，我们证明，HN亚基，这是通常负责病毒结合，似乎也是必不可少的融合活性，因为它的破坏消除溶血和融合，但不凝集，由缀合物修饰的病毒。
A method is described for preparing a covalent conjugate of proteins, in particular antibodies and their fragments, with gangliosides in the micellar form. The protein-ganglioside conjugate is associated with ganglioside micelles and can be separated from free protein by molecular sieve chromatography. Conjugates can irreversibly transfer from the micelle to a cell membrane of choice, and the protein portion be identified as a new surface antigen. The successful application of this methodology has been demonstrated with three biological systems. Rabbit IgG-ganglioside conjugate has been transferred to human or sheep erythrocytes, which have been hemagglutinated with goat anti-rabbit IgG. Erythrocytes modified with ganglioside-anti-H2Kkhave been shown to adhere to monolayers of L929 mouse fibroblasts which express H2Kk-antigen. Mouse monoclonal anti-glycophorin ganglioside conjugate can associate with Sendai virus and confer upon the virus the ability to agglutinate and hemolyse desialylated human erythrocytes. Using the anti-glycophorin conjugate, we demonstrated that the HN subunit, which is normally responsible for viral binding, appears also to be essential for fusion activity, because its destruction eliminates hemolysis and fusion, but not agglutination, by the conjugate-modified virus.