TAT fusion protein transduction into isolated mitochondria is accelerated by sodium channel inhibitors.
TAT fusion protein transduction into isolated mitochondria is accelerated by sodium channel inhibitors.
复制标题
DOI：
10.1021/bi101057v
复制
发表时间：
2010-11-09
期刊：
影响因子：
2.9
通讯作者：
Payne, R. Mark
中科院分区：
文献类型：
--
作者：
Rayapureddi, Jayanagendra P.;Tomamichel, Wendy J.;Walton, Sonia T.;Payne, R. Mark
严格控制离子和蛋白质在线粒体膜上的运输是维持线粒体功能和生物发生所必需的。特别是，线粒体内膜通常不允许蛋白质进入基质，除非通过严格调节的蛋白质进口机制。最近，细胞渗透肽已被证明以一种独立的机制穿过线粒体内膜。HIV-1转录反激活因子（TAT）是一种富含精氨酸的细胞渗透肽，47YGRKKRRQRRR57，它不仅可以转导全长蛋白穿过细胞膜，还可以转导到胞内细胞器。在这项研究中，我们研究了含有tat的蛋白质进入线粒体基质的能力。通过对分离纯化的线粒体进行新的FACS分析，我们发现TAT可以将修饰的荧光蛋白mMDH-GFP传递到线粒体基质中，并随后被基质肽酶处理。此外，TAT-mMDH-GFP进入线粒体的转导不依赖于典型蛋白的输入途径以及线粒体膜电位。与已发表的关于细胞膜的报道形成鲜明对比的是，钠通道抑制剂阿米洛利（amiloride）可阻断胞吞作用并抑制TAT转导，而同样的钠通道抑制剂可显著增加TAT转导到线粒体。这些结果证实，细胞渗透肽TAT可以很容易地将蛋白质货物转导到线粒体基质中。这些结果也证明了线粒体钠通道在介导TAT转导到线粒体中独立于内吞机制的新作用。因此，TAT转导到线粒体的机制与跨细胞膜的转导明显不同。
Stringent control of ion and protein transport across the mitochondrial membranes is required to maintain mitochondrial function and biogenesis. In particular, the inner mitochondrial membrane is generally impermeable to proteins entering the matrix except via tightly regulated protein import mechanisms. Recently, cell penetrant peptides have been shown to move across the inner mitochondrial membrane in a manner suggesting an independent mechanism. HIV-1 transactivator of transcription (TAT) is an arginine-rich cell penetrant peptide, 47YGRKKRRQRRR57, which can transduce full-length proteins not only across the cell membrane but also into intracellular organelles. In this study, we investigated the ability of a TAT-containing protein to move into the mitochondrial matrix. Using a novel FACS assay for isolated, purified mitochondria, we show that TAT can deliver a modified fluorescent protein, mMDH-GFP, to the matrix of mitochondria and it is subsequently processed by the matrix peptidases. In addition, transduction of TAT-mMDH-GFP into mitochondria is independent of canonical protein import pathways as well as mitochondrial membrane potential. In direct contrast to published reports regarding the cell membrane where the sodium channel inhibitor, amiloride, blocks endocytosis and inhibits TAT transduction, TAT transduction into mitochondria is markedly increased by this same sodium channel inhibitor. These results confirm that the cell penetrant peptide, TAT, can readily transduce a protein cargo into the mitochondrial matrix. These results also demonstrate a novel role for mitochondrial sodium channels in mediating TAT transduction into mitochondria that is independent of endocytotic mechanisms. The mechanism of TAT transduction into mitochondria therefore is distinctly different from transduction across the cell membrane.
登录
查看更多内容
影响因子：
3.9
作者：
Futaki, S.;Nakase, I.;Jones, A. T.
通讯作者：
Jones, A. T.
影响因子：
3.3
作者：
Garciarena, Carolina D.;Caldiz, Claudia I.;Ennis, Irene L.
通讯作者：
Ennis, Irene L.
影响因子：
4.8
作者：
Eguchi, A;Akuta, T;Nakanishi, M
通讯作者：
Nakanishi, M
影响因子：
3.5
作者：
Lim, MLR;Minamikawa, T;Nagley, P
通讯作者：
Nagley, P
影响因子：
12.4
作者：
Del Gaizo, V;Payne, RM
通讯作者：
Payne, RM