Reassembled HVJ (Sendai virus) envelopes containing non-toxic mutant proteins of diphtheria toxin show toxicity to mouse L cell
Reassembled HVJ (Sendai virus) envelopes containing non-toxic mutant proteins of diphtheria toxin show toxicity to mouse L cell
复制标题
含有白喉毒素无毒突变蛋白的重组 HVJ（仙台病毒）包膜对小鼠 L 细胞显示出毒性
作者：
T. Uchida;M. Yamaizumi;Y. Okada
该实验室的先前研究已经描述了从用非离子去污剂溶解的HVJ病毒体制备不含核衣壳的重组包膜部分。这样的重组包膜仍然保留神经氨酸酶、血细胞凝集、细胞融合和溶血活性1 -3。由于完整的病毒粒子在与哺乳动物细胞融合后将其核蛋白释放到细胞质中，因此似乎可以通过这种方式将包含在这种重新组装的病毒包膜内的任何给定的大分子引入易感的哺乳动物细胞中。我们在这里报告的成功应用这种方法引入CRM 45，一个无毒的突变蛋白相关的白喉毒素4，到小鼠L细胞，一个菌株，通常是毒素抗性。虽然CRM 45是一种有效的NAD：延伸因子2-ADP核糖基（NAD：EF 2-ADPR）转移酶5，6，因此阻断真核细胞提取物中的蛋白质合成，但它对动物细胞是无毒的，因为它缺乏与易感细胞膜结合所必需的氨基酸的C末端序列，因此不能到达细胞膜7。如果CRM 45的一些分子可以被引入活细胞的细胞质中，它们应该会阻止蛋白质合成并杀死细胞。这现在已经通过将含有CRM 45的重组HVJ包膜与培养的小鼠L细胞融合来实现。
PREVIOUS studies from this laboratory have described the preparation, from HVJ virions solubilised with a non-ionic detergent, of a reassembled envelope fraction free of nucleocapsids. Such reconstituted envelopes still retain neuraminidase, haemagglutinating, cell fusion and haemolytic activities1–3. Since intact virions release their nucleoproteins into the cytoplasm after fusion with mammalian cells, it seemed possible that any given macromolecule contained within such a reassembled viral envelope might be introduced into susceptible mammalian cells by this means. We report here the successful application of this method to the introduction of CRM45, a non-toxic mutant protein related to diphtheria toxin4, into mouse L cells, a strain that is normally toxin resistant. Although CRM45 is a potent NAD : elongation factor 2-ADP ribosyl (NAD : EF2-ADPR) transferase5,6 and thus blocks protein synthesis in eukaryotic cell extracts, it is non-toxic for animal cells because it lacks a C-terminal sequence of amino acids necessary to bind to susceptible cell membranes and thus cannot reach the cell interior7. If a few molecules of CRM45 could be introduced into the living cell cytoplasm, they should block protein synthesis and kill the cells. This has now been accomplished by fusion of CRM45-containing reassembled HVJ envelopes with cultured mouse L cells.