Development of small-molecular-bioprobes and proteomic analyses of signal transduction factors.

为了阐明生物功能，有两种分析方法。一种是基于信息和结构的分析，如基因组科学或结构生物学。另一种是基于化学生物学的分析，其中生物过程所必需的蛋白质使用其特定抑制剂进行鉴定。本研究项目的目的是产生对后一种研究有用的化学物质（“生物探针”），用于调节细胞中的信号转导途径，识别生物探针的生物靶分子，并挖掘和分析新的分子靶标。为此，我们开展了该项目，并取得了以下许多重要成果。(1)开发光交联小分子微阵列技术，以高通量方式鉴定靶蛋白的特异性相互作用分子。Angew.Chem.Int.Ed。(2)磷霉素与其特异性分子靶点蛋白磷酸酶2A的结合机制的阐明。2月。（2005）(3)鉴定线粒体特异性热休克蛋白Hsp60作为epolactaene的靶分子，并阐明抑制Hsp60伴侣活性的机制。Biochem.J。(2005)2。利用新型生物探针研究细胞信号转导通路(1)确定了一种放线菌生物合成PP2A特异性抑制剂光生菌素（phoslactomycin）的途径和基因簇，并开发了一种增强光生菌素在体内选择性生产的方法。J.Biol.Chem四面体(2003)。(2)新型血管生成抑制剂RK805和环氧双酚A的分离与表征。四面体(2004),Chem.Commun。(2005)3。(1)细胞周期调节蛋白激酶的泛素-蛋白酶体依赖性降解机制的阐明，Weel。Proc.Natl.Acad.Sci。美国(2004)更少

To elucidate biological functions, there are two ways of analysis. One is the information and structure based analysis such as genome science or structural biology. The other is the chemical biology based analysis, in which proteins essential for biological process are identified using their specific inhibitors. The purposes of the present research project are to generate chemicals ("bioprobes") useful for the latter type of study, which regulate the signal transduction pathway in cells, to identify biological target molecules of the bioprobes and to mine and analyze novel molecular targets. We have conducted the project for these purposes and have obtained many important results as follows.1.Identification of molecular targets and characterization of bioprobes using proteomic analysis(1)Development of photo-crosslinked small molecule microarrays to identify specific interacting molecules for target proteins in high throughput manner. Angew.Chem.Int.Ed.(2005)(2)Elucidation of the bindi … More ng mechanism of phoslactomycin to its specific molecular target, protein phosphatase 2A.FEBS Lett.(2005)(3)Identification of a mitochondria specific heat shock protein, Hsp60 as a target molecule for epolactaene and elucidation of the mechanism to inhibit HSP60's chaperone activity. Biochem.J.(2005)2.Exploitation of novel bioprobes for the study of cellular signal transduction pathway(1)Identification of the pathway and the gene cluster in an actinomycete for the biosynthesis of PP2A specific inhibitor, phoslactomycins, and development of a method to enhance the selective production of phoslactomycins in vivo. Tetrahedron (2003),J.Biol.Chem.(2003)(2)Isolation and characterization of novel compounds, RK805 and epoxytwinol A as novel angiogenesis inhibitors. Tetrahedron (2004),Chem.Commun.(2005)3.Molecular biology on novel molecular targets(1)Elucidation of the mechanism of ubiquitin-proteasome dependent degradation of a cell cycle regulatory protein kinase, Weel. Proc.Natl.Acad.Sci.USA(2004) Less