植物中的邻近标签。

IF 21.3 1区 生物学 Q1 PLANT SCIENCES Annual review of plant biology Pub Date : 2023-05-22 Epub Date: 2023-02-28 DOI:10.1146/annurev-arplant-070522-052132
Shou-Ling Xu, Ruben Shrestha, Sumudu S Karunadasa, Pei-Qiao Xie
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引用次数: 4

摘要

蛋白质是细胞中的主力;它们形成稳定的、更经常是动态的、短暂的蛋白质-蛋白质相互作用、组装和网络,并与DNA和RNA密切相互作用。这些网络相互作用是基本生物过程的基础,在细胞功能中发挥着重要作用。最近,与质谱法(PL-MS)相结合的邻近依赖性生物素标记方法已成为在分子水平上剖析复杂细胞网络的一种强大技术。在PL-MS中,通过将遗传编码的邻近标记(PL)酶融合到蛋白质或定位信号肽,该酶被靶向感兴趣的蛋白质复合物或细胞器,从而允许在缩放半径内标记邻近蛋白质。然后,这些生物素化的蛋白质可以被链亲和素珠捕获,并通过质谱法进行鉴定和定量。最近工程化的PL酶,如TurboID,具有显著提高的酶活性,能够以显著提高的信噪比进行时空映射。PL-MS通过克服传统技术带来的几个障碍,如生化分馏和亲和纯化质谱,彻底改变了我们进行蛋白质组学的方式。在这篇综述中,我们重点关注已经或可能被植物领域采用的基于生物素连接酶的PL-MS应用。我们讨论了实验设计,并回顾了工程化生物素连接酶的不同选择、富集和定量策略。最后,我们回顾了验证并讨论了未来的展望。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Proximity Labeling in Plants.

Proteins are workhorses in the cell; they form stable and more often dynamic, transient protein-protein interactions, assemblies, and networks and have an intimate interplay with DNA and RNA. These network interactions underlie fundamental biological processes and play essential roles in cellular function. The proximity-dependent biotinylation labeling approach combined with mass spectrometry (PL-MS) has recently emerged as a powerful technique to dissect the complex cellular network at the molecular level. In PL-MS, by fusing a genetically encoded proximity-labeling (PL) enzyme to a protein or a localization signal peptide, the enzyme is targeted to a protein complex of interest or to an organelle, allowing labeling of proximity proteins within a zoom radius. These biotinylated proteins can then be captured by streptavidin beads and identified and quantified by mass spectrometry. Recently engineered PL enzymes such as TurboID have a much-improved enzymatic activity, enabling spatiotemporal mapping with a dramatically increased signal-to-noise ratio. PL-MS has revolutionized the way we perform proteomics by overcoming several hurdles imposed by traditional technology, such as biochemical fractionation and affinity purification mass spectrometry. In this review, we focus on biotin ligase-based PL-MS applications that have been, or are likely to be, adopted by the plant field. We discuss the experimental designs and review the different choices for engineered biotin ligases, enrichment, and quantification strategies. Lastly, we review the validation and discuss future perspectives.

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来源期刊
Annual review of plant biology
Annual review of plant biology 生物-植物科学
CiteScore
40.40
自引率
0.40%
发文量
29
期刊介绍: The Annual Review of Plant Biology is a peer-reviewed scientific journal published by Annual Reviews. It has been in publication since 1950 and covers significant developments in the field of plant biology, including biochemistry and biosynthesis, genetics, genomics and molecular biology, cell differentiation, tissue, organ and whole plant events, acclimation and adaptation, and methods and model organisms. The current volume of this journal has been converted from gated to open access through Annual Reviews' Subscribe to Open program, with all articles published under a CC BY license.
期刊最新文献
Adaptation and the Geographic Spread of Crop Species. Environmental Control of Hypocotyl Elongation. Plant Cryopreservation: Principles, Applications, and Challenges of Banking Plant Diversity at Ultralow Temperatures. Structure and Function of Auxin Transporters. Structural and Evolutionary Aspects of Plant Endocytosis.
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