A comprehensive model for the cellular uptake of cationic cell-penetrating peptides
A comprehensive model for the cellular uptake of cationic cell-penetrating peptides
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DOI：
10.1111/j.1600-0854.2007.00572.x
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发表时间：
2007-07-01
期刊：
影响因子：
4.5
通讯作者：
Brock, Roland
中科院分区：
文献类型：
--
作者：
Duchardt, Falk;Fotin-Mleczek, Mariola;Brock, Roland
质膜代表了大多数大分子的不可渗透屏障。还有一些蛋白质和所谓的细胞穿透肽有效地进入细胞。已经表明，内吞作用有助于这些分子的输入。然而，关于内吞过程的性质，已经获得了相互矛盾的结果。此外，已经有了新的发现，一个内吞作用无关的细胞进入。在这项研究中，我们提供的证据表明，触角足同源结构域（Antp）肽，九精氨酸和HIV-1 Tat蛋白衍生的达特肽同时使用三种内吞途径：巨胞饮，网格蛋白介导的内吞和小窝/脂筏介导的内吞。在不同的输入机制对摄取的贡献程度上，Apnapedia与达特和R9不同。此外，在较高浓度下，通过源自质膜的空间限制位点的机制发生摄取，并导致肽的快速细胞质分布。不能检测到内吞囊泡，这表明一种不依赖于内吞作用的摄取模式。肝素酶处理的细胞负面影响这一进口，蛋白激酶C抑制剂rottlerin，显性负动力蛋白和氯丙嗪的表达。在一组不同的细胞系中观察到这种摄取机制。对于Antp，需要显著更高的肽浓度和抑制内吞作用来诱导其摄取。这些调查结果的相关性的生物活性货物的进口。
The plasma membrane represents an impermeable barrier for most macromolecules. Still some proteins and so-called cell-penetrating peptides enter cells efficiently. It has been shown that endocytosis contributes to the import of these molecules. However, conflicting results have been obtained concerning the nature of the endocytic process. In addition, there have been new findings for an endocytosis-independent cellular entry. In this study, we provide evidence that the Antennapedia-homeodomain-derived antennapedia (Antp) peptide, nona-arginine and the HIV-1 Tat-protein-derived Tat peptide simultaneously use three endocytic pathways: macropinocytosis, clathrin-mediated endocytosis and caveolae/lipid-raft-mediated endocytosis. Antennapedia differs from Tat and R9 by the extent by which the different import mechanisms contribute to uptake. Moreover, at higher concentrations, uptake occurs by a mechanism that originates from spatially restricted sites of the plasma membrane and leads to a rapid cytoplasmic distribution of the peptides. Endocytic vesicles could not be detected, suggesting an endocytosis-independent mode of uptake. Heparinase treatment of cells negatively affects this import, as does the protein kinase C inhibitor rottlerin, expression of dominant-negative dynamin and chlorpromazine. This mechanism of uptake was observed for a panel of different cell lines. For Antp, significantly higher peptide concentrations and inhibition of endocytosis were required to induce its uptake. The relevance of these findings for import of biologically active cargos is shown.