Membrane proteome in Escherichia coli probed by MS3 mass spectrometry:: a preliminary report
Membrane proteome in Escherichia coli probed by MS3 mass spectrometry:: a preliminary report
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DOI：
10.1002/rcm.3104
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发表时间：
2007-01-01
影响因子：
2
通讯作者：
Amoresano, Angela
中科院分区：
文献类型：
--
作者：
Cirulli, Claudia;Marino, Gennaro;Amoresano, Angela
膜蛋白质组的表征是特别有趣的，因为在这一领域的更好的知识可能会导致新的见解不同的膜系统的功能。尽管表面蛋白的生物相关性，然而，它们的表征仍然是一个具有挑战性的任务。革兰氏阴性菌的外膜蛋白（OMPs）是细胞与环境界面的关键分子。因此，革兰氏阴性菌的表面蛋白含有可能是药物、抗微生物剂或检测系统的良好靶标的蛋白质，并且它们可能成为有效疫苗的组分。在这方面，大肠杆菌已被选为模式生物的几个结构和功能的研究，旨在了解革兰氏阴性细胞壁中的蛋白质的生物物理和生物化学组织。在这里，我们提出了第一个结果，为鉴定细菌表面暴露的蛋白质在大肠杆菌。coli K12的基础上使用丹磺酰氯标记耦合二维串联质谱利用前体离子/MS 3扫描模式的优势。该方法为鉴定大肠杆菌膜蛋白提供了一种有前途的、简单的、快速的方法。大肠杆菌作为模式生物，从而避免了基于二维液相色谱和电泳的耗时程序。所鉴定的蛋白质可分为五个主要家族：外膜（29种蛋白质）、脂蛋白（6种蛋白质）、跨膜（43种蛋白质）家族。版权所有（C）2007约翰威利父子有限公司
Characterization of the membrane proteome is particularly intriguing since a better knowledge in this field might lead to new insights into the function of different membrane systems. Despite the biological relevance of surface proteins however, their characterization still remains a challenging task. Outer membrane proteins (OMPs) of Gram-negative bacteria are key molecules that interface the cell with the environment. Hence, surface proteins of Gram-negative bacteria contain proteins that might be good targets for drugs, antimicrobials or detection systems and they may become components of effective vaccines. In this respect, Escherichia coli has been chosen as a model organism for several structural and functional studies aimed at understanding the biophysical and biochemical organization of proteins in Gram-negative cell walls. Here we present first results for the identification of bacterial surface exposed proteins in E. coli K12 based on the use of dansyl chloride labelling coupled with bidimensional tandem mass spectrometry exploiting the advantage of precursor ion/MS3 scan modes. This procedure resulted in a promising, simple, and rapid strategy for the identification of membrane proteins in E. coli as model organism, thus avoiding time-consuming procedures based on two-dimensional liquid chromatography and electrophoresis. The proteins identified could be grouped into five major families: outer membrane (29 proteins), lipoproteins (6 proteins), transmembrane (43 proteins) families. Copyright (C) 2007 John Wiley & Sons, Ltd.