Oligoarginine-Bearing Tandem Repeat Penetration-Accelerating Sequence Delivers Protein to Cytosol via Caveolae-Mediated Endocytosis
Oligoarginine-Bearing Tandem Repeat Penetration-Accelerating Sequence Delivers Protein to Cytosol via Caveolae-Mediated Endocytosis
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DOI：
10.1021/acs.biomac.8b01299
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发表时间：
2019-05-01
影响因子：
6.2
通讯作者：
Futaki, Shiroh
中科院分区：
文献类型：
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作者：
Okuda, Akiko;Tahara, Shinya;Futaki, Shiroh
为了促进蛋白质等较大分子的胞质递送，我们开发了一种新的细胞穿透肽序列，名为 Pas2r12，由重复的 Pas 序列 (FFLIG-FFLIG) 和 D-十二精氨酸 (r12) 组成。该肽显着增强了作为货物的增强型绿色荧光蛋白和免疫球蛋白 G 的细胞摄取和胞质释放。我们发现，简单地将 Pas2r12 与货物混合就可以产生胞质可引入的形式。 Pas2r12 的胞质转运被发现是一个需要能量的过程，依赖于肌动蛋白聚合，并受到小凹介导的内吞作用抑制剂（染料木黄酮和甲基-β-环糊精）和针对小凹蛋白-1 的小干扰 RNA 的抑制。这些结果表明，Pas2r12 无需交联即可增强货物的膜渗透，并且小凹介导的内吞作用可能是增强胞质递送的途径。
To facilitate the cytosolic delivery of larger molecules such as proteins, we developed a new cell-penetrating peptide sequence, named Pas2r12, consisting of a repeated Pas sequence (FFLIG-FFLIG) and D-dodecaarginine (r12). This peptide significantly enhanced the cellular uptake and cytosolic release of enhanced green fluorescent protein and immunoglobulin G as cargos. We found that simply mixing Pas2r12 with cargos could generate cytosolic introducible forms. The cytosolic delivery of cargos by Pas2r12 was found to be an energy-requiring process, to rely on actin polymerization, and to be suppressed by caveolae-mediated endocytosis inhibitors (genistein and methyl-beta-cyclodextrin) and small interfering RNA against caveolin-1. These results suggest that Pas2r12 enhances membrane penetration of cargos without the need for cross-linking and that caveolae-mediated endocytosis may be the route by which cytosolic delivery is enhanced.