Zieske Lynn R
Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, USA.
J Exp Bot. 2006;57(7):1501-8. doi: 10.1093/jxb/erj168. Epub 2006 Mar 30.
Proteomic research includes the characterization of protein mixtures in order to understand complex biological systems and determine relationships between proteins, their function, and protein-protein interactions. Often the goal of such research is to monitor changes of proteins in perturbed systems, a type of study referred to as differential expression analysis. To perform these studies requires the ability to execute some type of differential comparison of a given protein state in reference to some type of a control. The iTRAQ reagents are a set of isobaric reagents which are amine specific and allow for the identification and quantitation of up to four different samples simultaneously. The amine specificity of these reagents makes most peptides in a sample amenable to this labeling strategy with no loss of information from samples involving post-translational modifications, such as the scrutiny of signal transduction pathways that often involve phosphorylation phenomena. In addition, the multiplexing capacity of these reagents allows for information replication within certain LC-MS/MS experimental regimes, providing additional statistical validation within any given experiment. The results presented herein demonstrate a few examples of the wide variety of quantitative information that can be realized when undertaking such experimental approaches. These include temporal analysis of drug-induced-protein expression, discovery and elucidation of disease markers, and protein-protein interactions in multi-protein complexes.
蛋白质组学研究包括对蛋白质混合物进行表征,以便理解复杂的生物系统,并确定蛋白质之间的关系、它们的功能以及蛋白质 - 蛋白质相互作用。此类研究的目标通常是监测受干扰系统中蛋白质的变化,这种研究类型称为差异表达分析。要进行这些研究,需要有能力针对某种类型的对照对给定蛋白质状态进行某种类型的差异比较。iTRAQ试剂是一组等压试剂,它们对胺具有特异性,能够同时鉴定和定量多达四个不同的样品。这些试剂的胺特异性使得样品中的大多数肽都适用于这种标记策略,而不会因涉及翻译后修饰的样品(例如经常涉及磷酸化现象的信号转导途径的详细研究)而丢失信息。此外,这些试剂的多重分析能力允许在某些液相色谱 - 串联质谱(LC-MS/MS)实验方案中进行信息复制,在任何给定实验中提供额外的统计验证。本文给出的结果展示了一些实例,说明采用此类实验方法时可实现的广泛定量信息。这些信息包括药物诱导的蛋白质表达的时间分析、疾病标志物的发现与阐释以及多蛋白复合物中的蛋白质 - 蛋白质相互作用。