Theodor-Echtermeyer-Weg 1, Leibniz-Institut für Gemüse- und Zierpflanzenbau, 14979 Groβbeeren, Germany.
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany.
Int J Mol Sci. 2021 Jul 1;22(13):7101. doi: 10.3390/ijms22137101.
The study of protein-protein interactions (PPIs) is fundamental in understanding the unique role of proteins within cells and their contribution to complex biological systems. While the toolkit to study PPIs has grown immensely in mammalian and unicellular eukaryote systems over recent years, application of these techniques in plants remains under-utilized. Affinity purification coupled to mass spectrometry (AP-MS) and proximity labeling coupled to mass spectrometry (PL-MS) are two powerful techniques that have significantly enhanced our understanding of PPIs. Relying on the specific binding properties of a protein to an immobilized ligand, AP is a fast, sensitive and targeted approach used to detect interactions between bait (protein of interest) and prey (interacting partners) under near-physiological conditions. Similarly, PL, which utilizes the close proximity of proteins to identify potential interacting partners, has the ability to detect transient or hydrophobic interactions under native conditions. Combined, these techniques have the potential to reveal an unprecedented spatial and temporal protein interaction network that better understands biological processes relevant to many fields of interest. In this review, we summarize the advantages and disadvantages of two increasingly common PPI determination techniques: AP-MS and PL-MS and discuss their important application to plant systems.
蛋白质-蛋白质相互作用（PPIs）的研究对于理解蛋白质在细胞中的独特作用及其对复杂生物系统的贡献至关重要。近年来，在哺乳动物和单细胞真核生物系统中，研究 PPIs 的工具包得到了极大的发展，但在植物中这些技术的应用仍未得到充分利用。亲和纯化结合质谱（AP-MS）和邻近标记结合质谱（PL-MS）是两种强大的技术，它们显著提高了我们对 PPIs 的理解。AP 依赖于蛋白质与固定化配体的特异性结合特性，是一种快速、敏感和靶向的方法，用于在接近生理条件下检测诱饵（感兴趣的蛋白质）和猎物（相互作用的伙伴）之间的相互作用。同样，PL 利用蛋白质的近距离来识别潜在的相互作用伙伴，具有在天然条件下检测瞬时或疏水性相互作用的能力。这些技术的结合有可能揭示一个前所未有的空间和时间蛋白质相互作用网络，更好地理解与许多感兴趣领域相关的生物学过程。在这篇综述中，我们总结了两种越来越常见的 PPI 测定技术（AP-MS 和 PL-MS）的优缺点，并讨论了它们在植物系统中的重要应用。
