Rapid and Sensitive Photoaffinity Labeling of Drug Receptors with Immobilized Probes

由于基因组和蛋白质组之间没有严格的线性关系，蛋白质组学是解决蛋白质网络的理想方法。直到最近，蛋白质组学几乎是变性蛋白质的二维凝胶电泳图谱的同义词。然而，关于蛋白质功能的主要问题是它们如何与它们的伴侣相互作用。因此，基于亲和性的方法作为化学基因组学中基于功能的蛋白质分选方法越来越受到人们的重视，光亲和性方法通过共价键将配体与其特异性受体交联，从而能够直接识别靶蛋白和配体结合结构域。由于蛋白质组学是对蛋白质的广泛研究，因此很明显，需要常规的光亲和方法来增加整个分析步骤的通量。我们首先开发了将光反应性二氮杂环丙烯与各种生物配体连接的有效方法。该方法显著增加了制备重要生物配体如DNA、RNA、碳水化合物、肽和蛋白质的各种光反应性衍生物的合成程序。与DNA技术类似，蛋白质芯片和蛋白质阵列是一种以大规模并行方式筛选样品的有效设备。因此，我们第二次开发了一种快速和灵敏的光亲和装置，用于捕获固体基质上的配体-受体对。基于表面等离子体共振光谱的技术被开发用于配体-受体相互作用的实时分析。与此相反，光亲和装置提供了一种用于分离靶蛋白的快速方法，其使得能够进行高通量质谱分析以鉴定靶及其功能位点。

Since there is no strict linear relationship between genome and proteome, proteomics is desirable to address protein networks. Until recently, proteomics was almost synonymous with the mapping of denatured proteins by two-dimensional gel electrophoresis. However, the major question about protein functions is how they interact with their partners. Thus, affinity-based methods have become increasingly appreciated as the approach of chemical genomics for sorting proteins based on their functions.Photoaffinity methods introduces a cross-link between a ligand and its specific receptor for probing their affinity correlation through a covalent bond, which enables the direct identification of target proteins as well as ligand binding domains. ^<1)>Because the proteomics of is the extensive study of proteins, it is clear that conventional photoaffinity methods are required to increase in the throughput throughout the analytical steps. We firstly developed efficient methods for tethering photoreactive diazirine to various biological ligands. The method significantly increases the synthetic routine in the preparation of various photoreactive derivatives of important biological ligands such as DNA,RNA,carbohydretes, peptides, and proteins. Analogous to DNA technologies, protein chips and protein arrays serve an efficient device for the Screening of samples in a massively parallel fashion. Thus, we secondly developed a rapid and sensitive photoaffinity device for capturing ligand-receptor pair on a solid matrix. The technique based on surface plasmon resonance spectroscopy was developed for the real-time analysis of ligand-receptor interaction. In contrast to this, the photoaffinity device provides a rapid method for the isolation of target proteins, which enables the high-throughput mass spectrometric analysis for the identification of targets as well as their functional sites.