Studies on the internalization mechanism of cationic cell-penetrating peptides
Studies on the internalization mechanism of cationic cell-penetrating peptides
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DOI：
10.1074/jbc.m303938200
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发表时间：
2003-08-15
影响因子：
4.8
通讯作者：
Temsamani, J
中科院分区：
文献类型：
--
作者：
Drin, G;Cottin, S;Temsamani, J
大量的数据表明阳离子肽能够以非温度依赖性的方式易位到真核细胞中。尽管这些肽被广泛用于促进生物活性分子的细胞内递送，但这种细胞穿透活性发生的机制仍然不清楚。在这里，我们提出了一个在体外研究的细胞摄取的肽，最初来自protegrin（合成B肽载体），也已被证明可以提高药物的运输通过血脑屏障。同时，我们研究了两个脂质相互作用肽SynB 5和pAntp-（43-58）的内化过程，后者对应于Apneapedia同源结构域的易位片段。我们报告的时间和剂量依赖性的内在化的定量研究，并证明这些肽积累囊泡结构内。此外，我们已经研究了内吞途径在这一过程中的作用，使用各种代谢和内吞抑制剂。我们表明，这些肽的内化是一个温度和能量依赖性的过程，内体转运是一个关键组成部分的机制。总之，我们的结果表明SynB和pAntp-（43-58）肽通过吸附介导的内吞过程而不是温度非依赖性易位渗透到细胞中。
A great deal of data has been amassed suggesting that cationic peptides are able to translocate into eucaryotic cells in a temperature-independent manner. Although such peptides are widely used to promote the intracellular delivery of bioactive molecules, the mechanism by which this cell-penetrating activity occurs still remains unclear. Here, we present an in vitro study of the cellular uptake of peptides, originally deriving from protegrin (the SynB peptide vectors), that have also been shown to enhance the transport of drugs across the blood-brain barrier. In parallel, we have examined the internalization process of two lipid-interacting peptides, SynB5 and pAntp-(43-58), the latter corresponding to the translocating segment of the Antennapedia homeodomain. We report a quantitative study of the time- and dose-dependence of internalization and demonstrate that these peptides accumulate inside vesicular structures. Furthermore, we have examined the role of endocytotic pathways in this process using a variety of metabolic and endocytosis inhibitors. We show that the internalization of these peptides is a temperature- and energy-dependent process and that endosomal transport is a key component of the mechanism. Altogether, our results suggest that SynB and pAntp-(43-58) peptides penetrate into cells by an adsorptive-mediated endocytosis process rather than temperature- independent translocation.