Jensen O N, Larsen M R, Roepstorff P
Department of Molecular Biology, Odense University, Denmark.
Proteins. 1998;Suppl 2:74-89. doi: 10.1002/(sici)1097-0134(1998)33:2+<74::aid-prot9>3.3.co;2-2.
The entire genomic DNA sequences of a number of prokaryotic and eukaryotic species are now available and many more, including the human genome, will be completed in the near future. The state-of-life of a cell at any given time, however, is defined by its protein composition, i.e., its proteome. Gel electrophoresis, mass spectrometry, and bioinformatics will be important tools for protein and proteome analysis in the post-genome era. Protein identification from electrophoretic gels by mass spectrometric peptide mapping or peptide sequencing combined with sequence database searching is established and has been applied to numerous biological systems. We describe current strategies and selected applications in molecular and cell biology. The next challenges are detailed structure/function analyses, which include studying the molecular composition of multiprotein complexes and characterization of secondary modifications of proteins. The advantages and limitations of a number of mass spectrometry-based strategies designed for microcharacterization of low amounts of protein from electrophoretic gels are discussed and illustrated by examples.
现在已有许多原核生物和真核生物物种的完整基因组DNA序列，包括人类基因组在内的更多序列将在不久的将来完成。然而，细胞在任何给定时间的生命状态是由其蛋白质组成，即蛋白质组决定的。凝胶电泳、质谱分析和生物信息学将成为后基因组时代蛋白质和蛋白质组分析的重要工具。通过质谱肽图谱或肽测序结合序列数据库搜索从电泳凝胶中鉴定蛋白质的方法已经确立，并已应用于众多生物系统。我们描述了分子和细胞生物学中的当前策略及选定的应用。接下来的挑战是进行详细的结构/功能分析，包括研究多蛋白复合物的分子组成以及蛋白质二级修饰的表征。本文讨论了多种基于质谱的策略用于对电泳凝胶中少量蛋白质进行微表征的优缺点，并举例说明。
