Split-protein systems: beyond binary protein-protein interactions.
Split-protein systems: beyond binary protein-protein interactions.
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DOI：
10.1016/j.cbpa.2011.10.014
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发表时间：
2011-12
影响因子：
7.8
通讯作者：
Ghosh, Indraneel
中科院分区：
文献类型：
--
作者：
Shekhawat, Sujan S.;Ghosh, Indraneel
据估计，在人类相互作用组中存在650，000种蛋白质-蛋白质相互作用，这是决定生命的所有可能的大分子伙伴关系的子集。因此，有一个持续的需要，为发展的敏感和用户友好的方法，在复杂的环境中编目生物大分子和检测它们的相互作用，修改，和细胞的位置。这样的方法还允许建立正常和患病细胞状态之间的相互作用组的差异，并允许量化治疗干预的结果。一个有前途的方法去卷积的大分子伙伴关系的作用是分裂蛋白质重组，也称为蛋白质片段互补。这种方法依赖于蛋白质报告分子的适当片段化，例如绿色荧光蛋白或萤火虫荧光素酶，当其连接到可能的相互作用伴侣时，可以重新组装并恢复功能，从而确认伙伴关系。裂解蛋白质方法已被有效地用于检测蛋白质间的相互作用，在无细胞系统中，E。大肠杆菌、酵母、哺乳动物细胞、植物和活的动物。在此，我们提出了工程分裂蛋白质系统，允许快速检测三元蛋白质复合物，小分子抑制剂，以及各种大分子，包括核酸，聚（ADP）核糖，和铁硫簇的最新进展。我们还提出了进展，结合联合收割机分裂蛋白质系统与化学诱导剂的二聚化策略，允许调节正交分裂蛋白酶的活性，以及在确定酶抑制剂的援助。最后，我们讨论了自抑制策略，导致打开传感器以及未来的方向分裂蛋白质的方法，包括可能的治疗方法。
It has been estimated that 650,000 protein-protein interactions exist in the human interactome, a subset of all possible macromolecular partnerships that dictate life. Thus there is a continued need for the development of sensitive and user-friendly methods for cataloguing biomacromolecules in complex environments and for detecting their interactions, modifications, and cellular location. Such methods also allow for establishing differences in the interactome between a normal and diseased cellular state and for quantifying the outcome of therapeutic intervention. A promising approach for deconvoluting the role of macromolecular partnerships is split-protein reassembly, also called protein fragment complementation. This approach relies on the appropriate fragmentation of protein reporters, such as the green fluorescent protein or firefly luciferase, which when attached to possible interacting partners can reassemble and regain function, thereby confirming the partnership. Split-protein methods have been effectively utilized for detecting protein-protein interactions in cell-free systems, E. coli, yeast, mammalian cells, plants, and live animals. Herein, we present recent advances in engineering split-protein systems that allow for the rapid detection of ternary protein complexes, small molecule inhibitors, as well as a variety of macromolecules including nucleic acids, poly(ADP) ribose, and iron sulfur clusters. We also present advances that combine split-protein systems with chemical inducers of dimerization strategies that allow for regulating the activity of orthogonal split-proteases as well as aid in identifying enzyme inhibitors. Finally, we discuss autoinhibition strategies leading to turn-on sensors as well as future directions in split-protein methodology including possible therapeutic approaches.
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影响因子：
82.9
作者：
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影响因子：
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通讯作者：
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