Gel-based and gel-free quantitative proteomics approaches at a glance.
Gel-based and gel-free quantitative proteomics approaches at a glance.
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DOI：
10.1155/2012/494572
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Sergeant K
中科院分区：
文献类型：
--
作者：
Abdallah C;Dumas-Gaudot E;Renaut J;Sergeant K
双向凝胶电泳（2-DE）被广泛应用，仍然是蛋白质组学的首选方法，然而，普遍的2-DE相关的问题破坏了其作为蛋白质组研究中的主要分离技术的前景。因此，最先进的鸟枪技术正在慢慢地接管和利用质谱（MS）的快速扩展和进步，以提供一个新的无凝胶定量技术工具箱。当与MS结合时，鸟枪蛋白质组学管道可以为蛋白质的灵敏和高通量分析提供新的途径，从而实现高准确性的定量。尽管基于标记的方法，无论是化学的还是代谢的，由于多路复用能力而在定量蛋白质组学中得到普及，但这些方法并非没有缺点。新兴的无标记方法是标签独立的，适用于所有类型的样品。由于蛋白质提取困难、绿色组织中蛋白质丰度较高以及缺乏注释完整的基因组序列，定量蛋白质组学的挑战在植物中更为突出。本透视分析的目标是呈现可用的基于凝胶的和无凝胶的方法及其对植物的应用的优势和劣势之间的平衡。最新的趋势，肽分馏服从MS分析进行了讨论。
Two-dimensional gel electrophoresis (2-DE) is widely applied and remains the method of choice in proteomics; however, pervasive 2-DE-related concerns undermine its prospects as a dominant separation technique in proteome research. Consequently, the state-of-the-art shotgun techniques are slowly taking over and utilising the rapid expansion and advancement of mass spectrometry (MS) to provide a new toolbox of gel-free quantitative techniques. When coupled to MS, the shotgun proteomic pipeline can fuel new routes in sensitive and high-throughput profiling of proteins, leading to a high accuracy in quantification. Although label-based approaches, either chemical or metabolic, gained popularity in quantitative proteomics because of the multiplexing capacity, these approaches are not without drawbacks. The burgeoning label-free methods are tag independent and suitable for all kinds of samples. The challenges in quantitative proteomics are more prominent in plants due to difficulties in protein extraction, some protein abundance in green tissue, and the absence of well-annotated and completed genome sequences. The goal of this perspective assay is to present the balance between the strengths and weaknesses of the available gel-based and -free methods and their application to plants. The latest trends in peptide fractionation amenable to MS analysis are as well discussed.