[医药卫生]生物大分子相互作用.ppt

二、概论1、分子细胞生物学前沿进展与技术 The biomacromolecules in cell The interaction of biomacromolecules is the fundamental basis of any cellular function The focuses: Protein-protein interaction Protein-DNA interaction Protein-RNA interaction The importance of protein-protein interaction Protein-protein interactions refer to the association of protein molecules and the study of these associations from the perspective of biochemistry, signal transduction and networks. Protein-protein interactions are involved in almost every aspects of biological processes. Protein-protein interactions provide potential drug targets. The methods commonly used for studying protein-protein interaction Chemical crosslinking 化学交联 Tagged Fusion Proteins (Pull-down) 沉降实验 Co-immunoprecipitation (Co-IP) 免疫共沉淀 Yeast Two-hybrid (Y2H) 酵母双杂交 Fluorescence Resonace Energy Trasfer (FRET) 荧光共振能量转移 Docking 嵌合计算 Biacore (Surface Plasmon Resonance, SPR) 表面等离子共振 Chemical Crosslinking Pull-down GST pull-down, His pull-down Co-Immunoprecipitation Example: BAG2 was identified by Co-IP as a CHIP-binding protein Fluorescence Resonance Energy Transfer (FRET) GFP and other FPs (fluorescent proteins) FRET: principle FRET is a distance-dependent physical process by which energy is transferred nonradiatively from an excited molecular fluorophore (the donor) to another fluorophore (the acceptor) The donor emission spectrum must significantly overlap the excitation spectrum of the acceptor 供体发射波与受体激发波有重叠 The donor and acceptor fluorophores must be in favorable mutual orientation 合适的空间取向 The distance between the donor and acceptor fluorophores must be less than 100 ? 距离100 ? Detecting methods Monitor changes in donor fluorescence 检测供体荧光 Monitor changes in acceptor fluorescence 检测受体荧光 Simultaneously measure changes in both donor and acceptor fluorescence using spectral imaging 同时检测受体和供体荧光 Spectral imaging Photobleaching Photobleaching: an example FRET: biosensor Docking Give the 3D structure of a protein target, compounds