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Protein-Protein Interaction Mapping by 2C-BioID

机译：蛋白质 - 蛋白质相互作用由2C-生物映射

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摘要

Protein-protein interactions (PPIs) add an essential layer of complexity to the information encoded by the genome. Modulation of such interactions is a key feature of most, if not all, cellular activities and allows cells to respond rapidly to both internal and external signals and stimuli. In this respect, the development of the BioID assay to interrogate PPIs within a cellular context represents an important adjunct to the range of tools currently at researchers' disposal. To address some of its current limitations, we devised 2C-BioID, in which biotin ligase and the protein of interest remain as separate entities until induced to associate. This is accomplished using the well-established FKBP-FRB dimerization system (based on the rapamycin-induced binding of FK506 binding protein and FKBP12-rapamycin binding domain.). The design of 2C-BioID ensures that biotin ligase association with the protein of interest occurs only after addition of the rapamycin analogue AP21967. As such, 2C-BioID alleviates potential targeting issues and improves the ability to exclude false positives, thereby refining the specificity of BiolD-generated interac-tomes.
机译：蛋白质 - 蛋白质相互作用（PPI）向由基因组编码的信息添加一个重要的复杂性。这种相互作用的调制是大多数的关键特征，如果不是全部，蜂窝活动，并且允许细胞快速响应内部和外部信号和刺激。在这方面，在蜂窝环境中向询问PPI的BioID测定的发展代表了目前研究人员所处理的工具范围的重要辅助。为了解决其目前的一些限制，我们设计了2℃-Bioid，其中生物素连接酶和感兴趣的蛋白质仍然是单独的实体，直至诱导助理。这是使用良好的FKBP-FRB二聚化系统完成的（基于雷帕霉素诱导的FK506结合蛋白和FKBP12-雷帕霉素结合结构域的结合。）。 2C-生物的设计确保仅在加入雷帕霉素类似物AP21967之后仅发生与感兴趣蛋白质的生物素连接酶联系。因此，2C-Bioid减轻了潜在的靶向问题，并提高了排除误报的能力，从而改善了Biold产生的互动性互动的特异性。

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  • 来源
    《Current Protocols in Cell Biology》 |2019年第1期|共11页
  • 作者

    Alexandre Chojnowski; Hendrikje Werner; Matthew Cook;

  • 作者单位

    Developmental and Regenerative Biology Institute of Medical Biology Agency for Science Technology and Research (A*STAR) Singapore Singapore;

    Developmental and Regenerative Biology Institute of Medical Biology Agency for Science Technology and Research (A*STAR) Singapore Singapore;

    Developmental and Regenerative Biology Institute of Medical Biology Agency for Science Technology and Research (A*STAR) Singapore Singapore;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
  • 关键词

    2C-BioID; AP21967; BiolD; FKBP; FRB; PPI;

    机译：2C Bioid;AP21967;Biold;FKBP;FRB;PPI;

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