通过二维凝胶电泳和质谱法对粟酒裂殖酵母进行蛋白质组分析。

Proteome analysis of Schizosaccharomyces pombe by two-dimensional gel electrophoresis and mass spectrometry.

作者信息

Hwang Kyung-Hoon, Carapito Christine, Böhmer Susanne, Leize Emmanuelle, Van Dorsselaer Alain, Bernhardt Rita

机构信息

Universität des Saarlandes, FR 8.3 Biochemie, Saarbrücken, Germany.

出版信息

Proteomics. 2006 Jul;6(14):4115-29. doi: 10.1002/pmic.200500863.

Abstract

The fission yeast Schizosaccharomyces pombe (S. pombe) is a unicellular eukaryote and contains many genes and regulatory mechanisms that are close to those of mammals. In this study, we performed a global proteomic analysis of the fission yeast S. pombe wild type h(-S) L 972 proteome. More than 1,500 protein spots were visualized on silver stained 2-D gels in the 3-10 pI range with a high resolution and high reproducibility. Protein identification was carried out by MALDI-TOF-MS and/or nanoLC-MS/MS. Advantage of the complementarity of these two MS approaches was used to enhance the identification quality. So far, 364 proteins (representing 157 different proteins) have been identified. We report here the identification of 117 new proteins on our 2-D reference map of this yeast compared to the first reference map. Of these identified proteins, 40.1% were involved in metabolism. The present work provides a very useful tool for all studies relying on S. pombe as a model organism and is a considerable complement to the first reference map of S. pombe published recently by Sun and coworkers (Sun, N., Jang, J., Lee, S., Kim, S. et al.., Proteomics 2005, 5, 1574-1579).

摘要

裂殖酵母粟酒裂殖酵母(S. pombe)是一种单细胞真核生物,含有许多与哺乳动物相近的基因和调控机制。在本研究中,我们对裂殖酵母S. pombe野生型h(-S) L 972蛋白质组进行了全蛋白质组分析。在3-10的等电点范围内,银染二维凝胶上清晰可见1500多个蛋白质点,分辨率高且重复性好。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和/或纳升液相色谱-串联质谱(nanoLC-MS/MS)进行蛋白质鉴定。利用这两种质谱方法的互补优势来提高鉴定质量。到目前为止,已鉴定出364种蛋白质(代表157种不同蛋白质)。与第一张参考图谱相比,我们在此报告在该酵母的二维参考图谱上鉴定出117种新蛋白质。在这些已鉴定的蛋白质中,40.1%参与代谢。本研究为所有以S. pombe作为模式生物的研究提供了一个非常有用的工具,是对Sun及其同事最近发表的S. pombe第一张参考图谱(Sun, N., Jang, J., Lee, S., Kim, S.等人,《蛋白质组学》2005年,第5卷,第1574 - 1579页)的重要补充。

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