Morimoto Kyoko, van der Hoorn Renier A L
The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657 Japan.
The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK
Plant Cell Physiol. 2016 Mar;57(3):446-61. doi: 10.1093/pcp/pcw003. Epub 2016 Feb 12.
The active proteome dictates plant physiology. Yet, active proteins are difficult to predict based on transcript or protein levels, because protein activities are regulated post-translationally in their microenvironments. Over the past 10 years, activity-based protein profiling (ABPP) is increasingly used in plant science. ABPP monitors the activities of hundreds of plant proteins using tagged chemical probes that react with the active site of proteins in a mechanism-dependent manner. Since labeling is covalent and irreversible, labeled proteins can be detected and identified on protein gels and by mass spectrometry using tagged fluorophores and/or biotin. Here, we discuss general concepts, approaches and practical considerations of ABPP, before we summarize the discoveries made using 40 validated probes representing 14 chemotypes that can monitor the active state of >4,500 plant proteins. These discoveries and new opportunities indicate that this emerging functional proteomic technology is a powerful discovery tool that will have an increasing impact on plant science.
活性蛋白质组决定植物生理状态。然而，基于转录本或蛋白质水平很难预测活性蛋白，因为蛋白质活性在其微环境中受到翻译后调控。在过去十年中，基于活性的蛋白质谱分析（ABPP）在植物科学中的应用越来越广泛。ABPP使用带标签的化学探针监测数百种植物蛋白的活性，这些探针以依赖机制的方式与蛋白质的活性位点发生反应。由于标记是共价且不可逆的，因此可以在蛋白质凝胶上以及通过使用带标签的荧光团和/或生物素的质谱法检测和鉴定标记的蛋白质。在此，我们在总结使用代表14种化学类型的40种经过验证的探针所取得的发现之前，先讨论ABPP的一般概念、方法和实际注意事项，这些探针可监测超过4500种植物蛋白的活性状态。这些发现和新机遇表明，这种新兴的功能蛋白质组学技术是一种强大的发现工具，将对植物科学产生越来越大的影响。
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