Olaya-Abril Alfonso, Jiménez-Munguía Irene, Gómez-Gascón Lidia, Rodríguez-Ortega Manuel J
Departamento de Bioquímica y Biología Molecular, Universidad de Córdoba, Campus de Excelencia Internacional CeiA3, Córdoba, Spain; Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC), Córdoba, Spain.
Departamento de Bioquímica y Biología Molecular, Universidad de Córdoba, Campus de Excelencia Internacional CeiA3, Córdoba, Spain; Departamento de Sanidad Animal, Universidad de Córdoba, Córdoba, Spain.
J Proteomics. 2014 Jan 31;97:164-76. doi: 10.1016/j.jprot.2013.03.035. Epub 2013 Apr 26.
Surface proteins play a critical role in the interaction between cells and their environment, as they take part in processes like signaling, adhesion, transport, etc. In pathogenic microorganisms, they can also participate in virulence or cytotoxicity. As these proteins have the highest chances to be recognized by the immune system, they are often the targets for the discovery of new vaccines. In addition, they can serve for the development of serological-based tools to diagnose infectious diseases. First-generation proteomic strategies for the identification of surface proteins rely on the biochemical fractionation and/or enrichment of this group of molecules or organelles containing them. However, in the last years, a novel second-generation approach has been developed, consisting of the digestion of live, intact cells with proteases, so that surface-exposed moieties (i.e. the "surfome" of a cell) are "shaved" and analyzed by LC/MS/MS. Here we review such a strategy, firstly set up and developed in Gram-positive bacteria, and further applied to Gram-negative bacteria, unicellular fungi, and also pluricellular organisms. We also discuss the advantages and inconvenients of the approach, and the still unresolved question about the intriguing presence of proteins predicted as cytoplasmic in the surfomes. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.
表面蛋白在细胞与其环境之间的相互作用中起着关键作用，因为它们参与信号传导、黏附、运输等过程。在致病微生物中，它们还可参与毒力或细胞毒性作用。由于这些蛋白最有可能被免疫系统识别，它们常常是新型疫苗研发的靶点。此外，它们可用于开发基于血清学的传染病诊断工具。第一代用于鉴定表面蛋白的蛋白质组学策略依赖于对这组分子或含有它们的细胞器进行生化分级分离和/或富集。然而，在过去几年中，已开发出一种新型的第二代方法，即使用蛋白酶消化活的完整细胞，从而“刮下”细胞表面暴露的部分（即细胞的“表面组”），并通过液相色谱/串联质谱进行分析。在此，我们综述这样一种首先在革兰氏阳性菌中建立和开发、并进一步应用于革兰氏阴性菌、单细胞真菌以及多细胞生物的策略。我们还讨论了该方法的优缺点，以及关于表面组中预测为细胞质蛋白的有趣存在这一仍未解决的问题。本文是名为《微生物蛋白质组学趋势》的特刊的一部分。
