Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization
Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization
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DOI：
10.1263/jbb.99.95
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发表时间：
2005-02-01
影响因子：
2.8
通讯作者：
Yamada, H
中科院分区：
文献类型：
--
作者：
Futami, J;Kitazoe, M;Yamada, H
在后基因组时代，迫切需要开发蛋白质操作方法来分析活细胞中蛋白质的功能。为此，目前正在评估将功能蛋白递送到活细胞中的技术，作为引入转录活性 DNA 的替代方法。在这里，我们描述了一种将蛋白质有效转导到活细胞中的新方法，其中蛋白质通过有限的化学缀合用聚乙烯亚胺（PEI）简单地阳离子化。 PEI 阳离子化蛋白质似乎通过离子电荷相互作用粘附在细胞表面，然后以不依赖于受体和转运蛋白的方式内化到细胞中。由于PEI是一种具有高阳离子电荷密度的有机大分子，与PEI的有限耦合导致蛋白质被赋予足够的阳离子电荷，而不会导致其基本功能的严重下降。许多PEI阳离子化蛋白，如核糖核酸酶（RNase）、绿色荧光蛋白（GFP）和免疫球蛋白（IgG），可以有效地进入细胞并在细胞质中发挥作用。我们的结果表明，使用 PEI 的蛋白质阳离子化技术将有助于蛋白质转导技术的开发。
In the post-genomic era, there is pressing need for development of protein manipulation methodology to analyze functions of proteins in living cells. For this purpose, techniques to deliver functional proteins into living cells are currently being evaluated as alternative approaches to the introduction of transcriptionally active DNA. Here, we describe a novel method for efficient protein transduction into living cells in which a protein is simply cationized with polyethylenimine (PEI) by limited chemical conjugation. PEI-cationized proteins appear to adhere to the cell surface by ionic charge interaction and then internalize into cells in a receptor- and transporter-independent fashion. Since PEI is an organic macromolecule with a high cationic-charge density, limited coupling with PEI results in endowment of sufficient cationic charge to proteins without causing serious decline in their fundamental functions. A number of PEI-cationized proteins, such as ribonuclease (RNase), green fluorescent protein (GFP) and immunoglobulin (IgG), efficiently entered cells and functioned in the cytosol. Our results suggest that protein cationization techniques using PEI will be useful for the development of protein transduction technology.