Homma M, Ouchi M
J Virol. 1973 Dec;12(6):1457-65. doi: 10.1128/JVI.12.6.1457-1465.1973.
Polypeptides of egg-borne Sendai virus (egg Sendai), which is biologically active on the basis of criteria of the infectivity for L cells and of hemolytic and cell fusion activities, were compared by polyacrylamide gel electrophoresis with those of L cell-borne (L Sendai) and HeLa cell-borne Sendai (HeLa Sendai) viruses, which are judged biologically inactive by the above criteria. Densitometer profiles on the stained gels of egg Sendai resolved six polypeptides (virion protein [VP] 1 to VP6), in which VP2 and VP4 were identified as glycoproteins by PAS stain. Comparative electropherograms of both L Sendai and HeLa Sendai revealed that there were significantly larger amounts in the VP2 region of these viruses but VP4 was present only in greatly reduced amounts as compared to egg Sendai. It was also found that VP2 of L Sendai and HeLa Sendai consisted of two components, VP2a and VP2b, but the one of egg Sendai consisted of only VP2a. A mild trypsin treatment which converts both L Sendai and HeLa Sendai to a biologically active form selectively removed VP2b from these viruses and increased concomitantly the amounts of materials in the VP4 region. The same treatment of egg Sendai affected neither its biological activities nor its electropherogram. Consequently, gross polypeptide profiles on the stained gels of L Sendai and HeLa Sendai after trypsin treatment became favorably comparable to that of egg Sendai. Electrophoresis of labeled L Sendai and HeLa Sendai with a (3)H-amino acids mixture and (14)C-glucosamine resolved at least three glycoproteins, GP1, GP2, and GP3, each corresponding to VP2a, VP2b, and VP4, respectively. The trypsin treatment of these viruses removed almost all the radioactivity of GP2 and simultaneously increased the radioactive counts of GP3 and raised small amounts of rapidly moving heterogeneous glycoprotein, GP4. A possible relationship between the biological modification and the above characteristic polypeptide patterns of Sendai virus was discussed.
根据对L细胞的感染性以及溶血和细胞融合活性标准具有生物活性的卵源仙台病毒(卵仙台病毒)的多肽,通过聚丙烯酰胺凝胶电泳与L细胞源(L仙台病毒)和HeLa细胞源仙台病毒(HeLa仙台病毒)的多肽进行比较,而根据上述标准判断这两种病毒无生物活性。对卵仙台病毒染色凝胶的光密度计分析解析出六种多肽(病毒体蛋白[VP]1至VP6),其中VP2和VP4经PAS染色鉴定为糖蛋白。L仙台病毒和HeLa仙台病毒的比较电泳图谱显示,这些病毒的VP2区域含量明显更高,但与卵仙台病毒相比,VP4的含量大幅减少。还发现,L仙台病毒和HeLa仙台病毒的VP2由两个组分VP2a和VP2b组成,但卵仙台病毒的VP2仅由VP2a组成。温和的胰蛋白酶处理可将L仙台病毒和HeLa仙台病毒转化为生物活性形式,该处理选择性地从这些病毒中去除VP2b,并同时增加VP4区域的物质含量。对卵仙台病毒进行相同处理既不影响其生物活性,也不影响其电泳图谱。因此,胰蛋白酶处理后L仙台病毒和HeLa仙台病毒染色凝胶上的总多肽图谱与卵仙台病毒的图谱具有良好的可比性。用(3)H -氨基酸混合物和(14)C -葡糖胺对标记的L仙台病毒和HeLa仙台病毒进行电泳,解析出至少三种糖蛋白,即GP1、GP2和GP3,分别对应于VP2a、VP2b和VP4。对这些病毒进行胰蛋白酶处理几乎去除了GP2的所有放射性,同时增加了GP3的放射性计数,并产生了少量快速移动的异质性糖蛋白GP4。讨论了仙台病毒生物修饰与上述特征性多肽模式之间的可能关系。