Mass spectrometric-based approaches in quantitative proteomics
Mass spectrometric-based approaches in quantitative proteomics
复制标题
DOI：
10.1016/s1046-2023(02)00303-1
复制
发表时间：
2003-02-01
期刊：
影响因子：
4.8
通讯作者：
Mann, M
中科院分区：
文献类型：
--
作者：
Ong, SE;Foster, LJ;Mann, M
传统上，旨在研究蛋白质表达变化的实验总是遵循一小组蛋白质。这种集中的方法是必要的，因为有效分析大量蛋白质的工具根本不可用。大规模的定量蛋白质组学有望产生大量的数据，而这些数据在以前用经典方法测量需要几十年的时间。质谱法已经是一种成熟的蛋白质鉴定工具，最近的方法学发展表明，它也可以成功地应用于提取蛋白质丰度的定量数据。从4年前的第一份报告开始，已经开发了许多利用稳定同位素核掺入蛋白质和肽的方案。在这里，我们回顾了一些最常用的协议的好处和陷阱，专注于一个程序，现在正在广泛使用在我们的实验室，稳定同位素标记与细胞培养中的氨基酸（SILAC）。讨论了SILAC实验的基本理论、应用和数据分析。这些技术的新兴性质和技术发展的快速步伐使得预测该领域的方向变得困难，但我们推测SILAC将很快成为定量蛋白质组学的关键工具。(C)2003 Elsevier Science（美国）。All rights reserved.
Classically, experiments aimed at studying changes in protein expression have always followed a small set of proteins. This focused approach was necessary since tools to efficiently analyze large numbers of proteins were simply not available. Large-scale quantitative proteomics promises to produce reams of data that previously would have taken decades to measure with classical methods. Mass spectrometry is already a well-established protein identification tool and recent methodological developments indicate that it can also be successfully applied to extract quantitative data of protein abundance. From the first reports 4 years ago, numerous schemes to take advantage of stable isotope nuclei incorporation in proteins and peptides have been developed. Here we review the benefits and pitfalls of some of the most commonly used protocols, focusing on a procedure now being used extensively in our laboratory, stable isotope labeling with amino acids in cell culture (SILAC). The basic theory, application, and data analysis of a SILAC experiment are discussed. The emerging nature of these techniques and the rapid pace of technological development make forecasting the directions of the field difficult but we speculate that SILAC will soon be a key tool of quantitative proteomics. (C) 2003 Elsevier Science (USA). All rights reserved.