使用亲和纯化-质谱法绘制差异蛋白质-蛋白质相互作用网络的方案。
Protocol for mapping differential protein-protein interaction networks using affinity purification-mass spectrometry.
作者信息
Kaushal Prashant, Ummadi Manisha R, Jang Gwendolyn M, Delgado Yennifer, Makanani Sara K, Alba Kareem, Winters Declan M, Blanc Sophie F, Xu Jiewei, Polacco Benjamin, Zhou Yuan, Stevenson Erica, Eckhardt Manon, Zuliani-Alvarez Lorena, Kaake Robyn, Swaney Danielle L, Krogan Nevan J, Bouhaddou Mehdi
机构信息
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, USA; Institute for Quantitative and Computational Biosciences, University of California, Los Angeles, Los Angeles, CA, USA; Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA, USA.
Quantitative Biosciences Institute (QBI), University of California, San Francisco, San Francisco, CA, USA; QBI Coronavirus Research Group (QCRG), University of California, San Francisco, San Francisco, CA, USA; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA; Gladstone Institute of Data Science and Biotechnology, J. David Gladstone Institutes, San Francisco, CA, USA.
出版信息
STAR Protoc. 2024 Dec 20;5(4):103286. doi: 10.1016/j.xpro.2024.103286. Epub 2024 Nov 2.
Proteins congregate into complexes to perform diverse cellular functions. Protein complexes are remodeled by protein-coding mutations or cellular signaling changes, driving phenotypic outcomes in health and disease. We present an affinity purification-mass spectrometry (AP-MS) proteomics protocol to express affinity-tagged "bait" proteins in mammalian cells, identify and quantify purified protein interactors, and visualize differential protein-protein interaction networks between pairwise conditions. Our protocol possesses general applicability to various cell types and biological areas. For complete details on the use and execution of this protocol, please refer to Bouhaddou et al..
蛋白质聚集形成复合物以执行多种细胞功能。蛋白质复合物通过蛋白质编码突变或细胞信号变化进行重塑，从而在健康和疾病中驱动表型结果。我们提出了一种亲和纯化-质谱（AP-MS）蛋白质组学方案，用于在哺乳动物细胞中表达带有亲和标签的“诱饵”蛋白质，鉴定和定量纯化的蛋白质相互作用物，并可视化成对条件之间的差异蛋白质-蛋白质相互作用网络。我们的方案对各种细胞类型和生物学领域具有普遍适用性。有关此方案的使用和执行的完整详细信息，请参考布哈杜等人的研究。
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