Larsen M R, Roepstorff P
Department of Molecular Biology, University of Southern Denmark, Odense University.
Fresenius J Anal Chem. 2000 Mar-Apr;366(6-7):677-90. doi: 10.1007/s002160051562.
High-throughput DNA sequencing has resulted in increasing input in protein sequence databases. Today more than 20 genomes have been sequenced and many more will be completed in the near future, including the largest of them all, the human genome. Presently, sequence databases contain entries for more than 425.000 protein sequences. However, the cellular functions are determined by the set of proteins expressed in the cell--the proteome. Two-dimensional gel electrophoresis, mass spectrometry and bioinformatics have become important tools in correlating the proteome with the genome. The current dominant strategies for identification of proteins from gels based on peptide mass spectrometric fingerprinting and partial sequencing by mass spectrometry are described. After identification of the proteins the next challenge in proteome analysis is characterization of their post-translational modifications. The general problems associated with characterization of these directly from gel separated proteins are described and the current state of art for the determination of phosphorylation, glycosylation and proteolytic processing is illustrated.
高通量DNA测序使得蛋白质序列数据库的输入不断增加。如今，已有20多个基因组完成了测序，在不久的将来还会有更多基因组完成测序，其中包括最大的人类基因组。目前，序列数据库包含超过42.5万个蛋白质序列条目。然而，细胞功能是由细胞中表达的一组蛋白质——蛋白质组决定的。二维凝胶电泳、质谱分析和生物信息学已成为将蛋白质组与基因组关联起来的重要工具。本文描述了基于肽质量谱指纹图谱和质谱部分测序从凝胶中鉴定蛋白质的当前主要策略。蛋白质鉴定之后，蛋白质组分析的下一个挑战是对其翻译后修饰进行表征。文中描述了直接从凝胶分离的蛋白质中表征这些修饰所面临的一般问题，并阐述了测定磷酸化、糖基化和蛋白水解加工的当前技术水平。
