Efficient intracellular delivery of a protein and a low molecular weight substance via recombinant polyomavirus-like particles
Efficient intracellular delivery of a protein and a low molecular weight substance via recombinant polyomavirus-like particles
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DOI：
10.1074/jbc.m313612200
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发表时间：
2004-06-25
影响因子：
4.8
通讯作者：
Reiser, COA
中科院分区：
文献类型：
--
作者：
Abbing, A;Blaschke, UK;Reiser, COA
基于核心蛋白VP2与VP1五聚体的特异性相互作用，开发了一种锚定技术，实现了将蛋白质和低分子量药物等外来分子有效地包封到多瘤病毒样颗粒（衣壳）中。VP2的49个氨基酸片段作为锚定分子，表达为与绿色荧光蛋白（GFP）融合蛋白或与甲氨蝶呤（MTX）共价连接。装载后的衣壳在几个月内表现出规律的形态和稳定性。通过小鼠成纤维细胞检测GFP和VP1荧光，以及细胞内释放的MTX对白血病T细胞的抑细胞作用，证实了GFP和MTX在体外内化到细胞中。
Efficient encapsulation of foreign molecules like proteins and low molecular weight drugs into polyoma virus-like particles (capsoids) was achieved by the development of an anchoring technique based upon the specific interaction of the inner core protein VP2 with VP1 pentamers. A stretch of 49 amino acids of VP2 served as an anchor molecule, either expressed as a fusion protein with green fluorescent protein (GFP) or covalently linked to methotrexate (MTX). The loaded capsoids showed regular morphology and stability for several months. GFP and MTX were internalized into cells in vitro, as was demonstrated by the detection of GFP and VP1 fluorescence in mouse fibroblasts and the cytostatic effect of intracellularly released MTX on leukemia T cells.