Department of Chemistry, University of Reading, Reading, UK.
Proteomics. 2011 Feb;11(4):756-75. doi: 10.1002/pmic.201000426. Epub 2011 Jan 18.
Quantitation is an inherent requirement in comparative proteomics and there is no exception to this for plant proteomics. Quantitative proteomics has high demands on the experimental workflow, requiring a thorough design and often a complex multi-step structure. It has to include sufficient numbers of biological and technical replicates and methods that are able to facilitate a quantitative signal read-out. Quantitative plant proteomics in particular poses many additional challenges but because of the nature of plants it also offers some potential advantages. In general, analysis of plants has been less prominent in proteomics. Low protein concentration, difficulties in protein extraction, genome multiploidy, high Rubisco abundance in green tissue, and an absence of well-annotated and completed genome sequences are some of the main challenges in plant proteomics. However, the latter is now changing with several genomes emerging for model plants and crops such as potato, tomato, soybean, rice, maize and barley. This review discusses the current status in quantitative plant proteomics (MS-based and non-MS-based) and its challenges and potentials. Both relative and absolute quantitation methods in plant proteomics from DIGE to MS-based analysis after isotope labeling and label-free quantitation are described and illustrated by published studies. In particular, we describe plant-specific quantitative methods such as metabolic labeling methods that can take full advantage of plant metabolism and culture practices, and discuss other potential advantages and challenges that may arise from the unique properties of plants.
定量分析是比较蛋白质组学的固有要求,植物蛋白质组学也不例外。定量蛋白质组学对实验工作流程要求很高,需要进行彻底的设计,并且通常具有复杂的多步骤结构。它必须包括足够数量的生物学和技术重复以及能够促进定量信号读取的方法。特别是定量植物蛋白质组学带来了许多额外的挑战,但由于植物的性质,它也提供了一些潜在的优势。一般来说,蛋白质组学中对植物的分析不太突出。植物蛋白质组学的主要挑战包括蛋白质浓度低、蛋白质提取困难、基因组多倍性、绿色组织中Rubisco 含量高以及缺乏经过充分注释和完成的基因组序列。然而,随着一些模式植物和作物(如马铃薯、番茄、大豆、水稻、玉米和大麦)的基因组的出现,这种情况正在发生变化。本文综述了定量植物蛋白质组学(基于 MS 和非 MS)的现状及其挑战和潜力。描述了 DIGE 到基于 MS 的分析后同位素标记和无标记定量的植物蛋白质组学中的相对和绝对定量方法,并通过已发表的研究进行了说明和举例。特别是,我们描述了植物特有的定量方法,例如代谢标记方法,这些方法可以充分利用植物代谢和培养实践,并讨论了可能由于植物独特性质而产生的其他潜在优势和挑战。