Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Department of Physiology and Cell Biology, Corrensstrasse 3, D-06466 Gatersleben, Germany.
Phytochemistry. 2011 Jul;72(10):963-74. doi: 10.1016/j.phytochem.2010.11.009. Epub 2010 Dec 20.
Recent innovations in liquid chromatography-mass spectrometry (LC-MS)-based methods have facilitated quantitative and functional proteomic analyses of large numbers of proteins derived from complex samples without any need for protein or peptide labelling. Regardless of its great potential, the application of these proteomics techniques to plant science started only recently. Here we present an overview of label-free quantitative proteomics features and their employment for analysing plants. Recent methods used for quantitative protein analyses by MS techniques are summarized and major challenges associated with label-free LC-MS-based approaches, including sample preparation, peptide separation, quantification and kinetic studies, are discussed. Database search algorithms and specific aspects regarding protein identification of non-sequenced organisms are also addressed. So far, label-free LC-MS in plant science has been used to establish cellular or subcellular proteome maps, characterize plant-pathogen interactions or stress defence reactions, and for profiling protein patterns during developmental processes. Improvements in both, analytical platforms (separation technology and bioinformatics/statistical analysis) and high throughput nucleotide sequencing technologies will enhance the power of this method.
近年来,基于液相色谱-质谱联用(LC-MS)的方法的创新使得无需对蛋白质或肽进行标记,就可以对来源于复杂样本的大量蛋白质进行定量和功能蛋白质组学分析。尽管具有巨大的潜力,但这些蛋白质组学技术在植物科学中的应用直到最近才开始。本文概述了无标记定量蛋白质组学的特点及其在植物分析中的应用。对 MS 技术定量蛋白质分析的最新方法进行了总结,并讨论了基于无标记 LC-MS 方法的主要挑战,包括样品制备、肽分离、定量和动力学研究。还讨论了数据库搜索算法以及关于非测序生物的蛋白质鉴定的具体方面。迄今为止,植物科学中的无标记 LC-MS 已用于建立细胞或亚细胞蛋白质组图谱、描述植物-病原体相互作用或应激防御反应,以及在发育过程中分析蛋白质图谱。分析平台(分离技术和生物信息学/统计分析)和高通量核苷酸测序技术的改进将增强该方法的功能。