Laboratorio de Proteómica, Centro de Investigación Príncipe Felipe, Avda, Autopista del Saler 16, 46012 Valencia, Spain.
J Proteomics. 2011 Feb 1;74(2):137-50. doi: 10.1016/j.jprot.2010.11.002. Epub 2010 Nov 11.
Proteomics is an expanding technology with potential applications in many research fields. Even though many research groups do not have direct access to its main analytical technique, mass spectrometry, they can interact with proteomics core facilities to incorporate this technology into their projects. Protein identification is the analysis most frequently performed in core facilities and is, probably, the most robust procedure. Here we discuss a few chemical reactions that are easily implemented within the conventional protein identification workflow. Chemical modification of proteins with N-hydroxysuccinimide esters, 4-sulfophenyl isothiocyanate, O-methylisourea or through β-elimination/Michael addition can be easily performed in any laboratory. The reactions are quite specific with almost no side reactions. These chemical tools increase considerably the number of applications and have been applied to characterize protein-protein interactions, to determine the N-terminal residues of proteins, to identify proteins with non-sequenced genomes or to locate phosphorylated and O-glycosylated.
蛋白质组学是一项不断发展的技术,具有广泛的应用前景。尽管许多研究小组无法直接使用其主要分析技术——质谱法,但它们可以与蛋白质组学核心设施合作,将该技术纳入其项目中。蛋白质鉴定是核心设施中最常进行的分析,可能也是最可靠的程序。在这里,我们讨论了一些在常规蛋白质鉴定工作流程中易于实施的化学反应。通过 N-羟基琥珀酰亚胺酯、4-磺基苯基异硫氰酸酯、O-甲基异脲或通过β-消除/迈克尔加成对蛋白质进行化学修饰,可以在任何实验室中轻松进行。这些反应具有很高的特异性,几乎没有副反应。这些化学工具大大增加了应用的数量,并已被应用于蛋白质-蛋白质相互作用的表征、确定蛋白质的 N 末端残基、鉴定无测序基因组的蛋白质以及定位磷酸化和 O-糖基化的蛋白质。