OLIGOPEPTIDES THAT SPECIFICALLY INHIBIT MEMBRANE-FUSION BY PARAMYXOVIRUSES - STUDIES ON THE SITE OF ACTION
OLIGOPEPTIDES THAT SPECIFICALLY INHIBIT MEMBRANE-FUSION BY PARAMYXOVIRUSES - STUDIES ON THE SITE OF ACTION
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DOI：
10.1016/0042-6822(83)90517-2
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发表时间：
1983-01-01
期刊：
影响因子：
3.7
通讯作者：
CHOPPIN, PW
中科院分区：
文献类型：
--
作者：
RICHARDSON, CD;CHOPPIN, PW
本实验室前期研究表明，氨基酸序列与副粘病毒F1多肽n端区序列相似的寡肽可抑制F蛋白的膜融合活性，从而在渗透水平上抑制病毒的感染性，并抑制病毒诱导的细胞融合和溶血。研究了这些寡肽抑制剂的作用位点。放射性标记的寡肽被发现能与细胞结合，但不能与病毒结合。4度预处理[非洲绿猴肾CV-1]细胞，而非病毒。C寡肽抑制麻疹病毒引起的感染起始和溶血。寡肽与细胞的结合在25度时是可逆的。或37.度。C.用氯甲基酮基合成寡肽，使它们能够不可逆结合，或者用叠氮基合成寡肽，使它们能够通过光激活在原位交联。这些衍生的寡肽保留了它们的抑制活性，结果证实它们与细胞结合，并在细胞上表达它们的抑制活性，而不是在病毒上。结果表明，寡肽与细胞膜上的受体位点发生反应，并通过与F1多肽竞争这些位点来抑制膜融合活性。一种寡肽与CV-1细胞结合的Scatchard分析显示，其结合的解离常数为1.2倍。10-7米，大约有。3.0的同学们。每个细胞有106个结合位点。
Previous studies from this laboratory showed that oligopeptides with amino acid sequences similar to the sequence of the N-terminal region of the F1 polypeptide of paramyxoviruses inhibited the membrane fusing activity of the F protein, and thereby inhibited virus infectivity at the level of penetration and virus-induced cell fusion and hemolysis. The site of action of these oligopeptide inhibitors has been investigated. Radioactively labeled oligopeptides were found to bind to cells, but not to virus. Pretreatment of [African green monkey kidney CV-1] cells, but not virus, at 4.degree. C with oligopeptides inhibited the initiation of infection and hemolysis induced by measles virus. The binding of the oligopeptides to cells was reversible at 25.degree. or 37.degree. C. Oligopeptides were synthesized with a chloromethylketone group to enable them to bind irreversibly, or with an azido group to permit them to be cross-linked in situ by photoactivation. The results with these derivatized oligopeptides, which retained their inhibitory activity, confirmed that they bind to, and express their inhibitory activity on, cells and not virus. The results suggest that the oligopeptides react with receptor sites on the cell membrane and inhibit membrane-fusing activity by competing with the F1 polypeptide for such sites. A Scatchard analysis of the binding of an oligopeptide to CV-1 cells revealed that it bound with a dissociation constant of 1.2 .times. 10-7 M and that there were .apprx. 3.0 .times. 106 binding sites per cell.