用于解旋酶功能表征的蛋白质-蛋白质相互作用分析
Protein-protein interaction analysis for functional characterization of helicases.
作者信息
Zybailov Boris L, Byrd Alicia K, Glazko Galina V, Rahmatallah Yasir, Raney Kevin D
机构信息
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, United States.
Department of Medical Informatics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, United States.
出版信息
Methods. 2016 Oct 1;108:56-64. doi: 10.1016/j.ymeth.2016.04.014. Epub 2016 Apr 20.
Helicases are enzymes involved in nucleic acid metabolism, playing major roles in replication, transcription, and repair. Defining helicases oligomerization state and transient and persistent protein interactions is essential for understanding of their function. In this article we review current methods for the protein-protein interaction analysis, and discuss examples of its application to the study of helicases: Pif1 and DDX3. Proteomics methods are our main focus - affinity pull-downs and chemical cross-linking followed by mass spectrometry. We review advantages and limitations of these methods and provide general guidelines for their implementation in the functional analysis of helicases.
解旋酶是参与核酸代谢的酶，在复制、转录和修复过程中发挥着重要作用。确定解旋酶的寡聚化状态以及瞬时和持久的蛋白质相互作用对于理解其功能至关重要。在本文中，我们综述了目前用于蛋白质-蛋白质相互作用分析的方法，并讨论了其在解旋酶研究中的应用实例：Pif1和DDX3。蛋白质组学方法是我们的主要关注点——亲和下拉法和化学交联后进行质谱分析。我们综述了这些方法的优缺点，并为其在解旋酶功能分析中的应用提供了一般指导原则。
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