Combinatorial targeting and discovery of ligand-receptors in organelles of mammalian cells.
Combinatorial targeting and discovery of ligand-receptors in organelles of mammalian cells.
复制标题
DOI：
10.1038/ncomms1773
复制
发表时间：
2012-04-17
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
噬菌体展示筛选允许研究细胞表面功能蛋白-蛋白相互作用，但研究细胞内细胞器仍然是一个挑战。在这里，我们引入内化噬菌体文库来鉴定通过受体独立机制进入哺乳动物细胞的克隆和靶向特异性细胞器，作为选择配体肽和鉴定其细胞内受体的工具。我们证明了穿透素，一种源自触角的肽，可以在噬菌体包膜上显示，并介导内化噬菌体进入细胞的非受体摄取。我们还发现，一个内化噬菌体结构显示了一个已建立的线粒体特异性定位信号，目标是线粒体，而一个内化噬菌体随机肽文库选择了定位于不同细胞内区室的肽基序。作为概念证明，我们证明了一种这样的肽，如果化学融合到穿透素，是内在化的受体独立，定位到线粒体，并促进细胞死亡。这种组合平台技术在细胞生物学和药物开发方面具有潜在的应用前景。噬菌体展示筛选可以揭示蛋白质与蛋白质之间的相互作用，但其应用主要局限于细胞表面。本文介绍了一种基于噬菌体的试剂，它可以将组合肽靶向细胞器，为发现细胞内配体受体提供了一种工具。
Phage display screening allows the study of functional protein–protein interactions at the cell surface, but investigating intracellular organelles remains a challenge. Here we introduce internalizing-phage libraries to identify clones that enter mammalian cells through a receptor-independent mechanism and target-specific organelles as a tool to select ligand peptides and identify their intracellular receptors. We demonstrate that penetratin, an antennapedia-derived peptide, can be displayed on the phage envelope and mediate receptor-independent uptake of internalizing phage into cells. We also show that an internalizing-phage construct displaying an established mitochondria-specific localization signal targets mitochondria, and that an internalizing-phage random peptide library selects for peptide motifs that localize to different intracellular compartments. As a proof-of-concept, we demonstrate that one such peptide, if chemically fused to penetratin, is internalized receptor-independently, localizes to mitochondria, and promotes cell death. This combinatorial platform technology has potential applications in cell biology and drug development. Phage display screening can unravel protein–protein interactions, but its application has been mainly restricted to the cell surface. Here, a phage-based reagent is introduced that allows the targeting of combinatorial peptides to cell organelles, providing a tool for the discovery of intracellular ligand-receptors.
登录
查看更多内容
影响因子：
4.5
作者：
Fabani, Martin M.;Gait, Michael J.
通讯作者：
Gait, Michael J.
影响因子：
5.3
作者：
Davidson, TJ;Harel, S;Troy, CM
通讯作者：
Troy, CM
影响因子：
4.8
作者：
Derossi, D;Calvet, S;Prochiantz, A
通讯作者：
Prochiantz, A
影响因子：
4.8
作者：
Eguchi, A;Akuta, T;Nakanishi, M
通讯作者：
Nakanishi, M
影响因子：
19
作者：
Derossi, D;Chassaing, G;Prochiantz, A
通讯作者：
Prochiantz, A