Cellular uptake of arginine-rich peptides: Roles for macropinocytosis and actin rearrangement
Cellular uptake of arginine-rich peptides: Roles for macropinocytosis and actin rearrangement
复制标题
DOI：
10.1016/j.ymthe.2004.08.010
复制
发表时间：
2004-12-01
影响因子：
12.4
通讯作者：
Futaki, S
中科院分区：
文献类型：
--
作者：
Nakase, I;Niwa, M;Futaki, S
使用膜渗透性肽作为用于细胞内递送各种蛋白质和大分子以改变细胞功能的载体载体已被充分记载。富含精氨酸的肽，包括衍生自人免疫缺陷病毒1达特蛋白的那些，是这些载体的代表性类别。这些载体肽及其蛋白缀合物的内化机制以前被认为与内吞作用分开，但最近的重新评估得出结论，内吞作用参与其内化。在这份报告中，我们表明，八精氨酸（R8）的HeLa细胞肽的摄取显着抑制巨胞饮抑制剂乙基异丙基amiloride（EIPA）和F-肌动蛋白聚合抑制剂细胞松弛素D，这表明巨胞饮在肽的摄取的作用。与此一致，我们观察到用R8肽处理细胞诱导肌动蛋白细胞骨架的显著重排。还观察到巨胞饮的内化效率和贡献对寡精氨酸肽的链长具有依赖性。另一种代表性的肽载体穿透素的摄取对EIPA不太敏感，并且穿透素对肌动蛋白定位没有如此明显的影响。上述观察结果表明，穿透素和R8肽具有不同的内化机制。
The use of membrane-permeable peptides as carrier vectors for the intracellular delivery of various proteins and macromolecules for modifying cellular function is well documented. Arginine-rich peptieds, including those derived from human immunodeficiency virus 1 Tat protein, are among the representative classes of these vectors. The internalization mechanism of these vector peptides and their protein conjugates was previously regarded as separate from endocytosis, but more recent reevaluations have concluded that endocytosis is involved in their internalization. In this report, we show that the uptake of octa-arginine (R8) peptide by HeLa cells was significantly suppressed by the macropinocytosis inhibitor ethylisopropylamiloride (EIPA) and the F-actin polymerization inhibitor cytochalasin D, suggesting a role for macropinocytosis in the uptake of the peptide. In agreement with this we observed that treatment of the cells with R8 peptide induced significant rearrangement of the actin cytoskeleton. The internalization efficiency and contribution of macropinocytosis were also observed to have a dependency on the chain length of the oligoarginine peptides. Uptake of penetratin, another representative peptide carrier, was less sensitive to EIPA and penetratin did not have such distinct effects on actin localization. The above observations suggest that penetratin and R8 peptides have distinct internalization mechanisms.