BIACORE 2000

这项共享仪器拨款提案的目的是为斯克里普斯研究所血管生物学系的七名研究人员购买一台生物传感器BIAcore 2000仪器。这些研究人员正在研究蛋白酶和蛋白酶抑制剂在血管系统中的结构和功能，整合素的结构和功能，载脂蛋白的结构和功能，激素原的加工和基因调控。该组研究者的研究领域与生理和病理过程相关，包括血栓溶解、动脉粥样硬化、心肌梗死、肥胖、癌症、高血压、神经元功能、发育和伤口愈合。这些研究强调蛋白质-蛋白质和蛋白质-DNA相互作用。由于这些研究中有几项是研究相同或相关的分子，因此我们将能够借鉴用户组其他成员的经验，设计在BIAcore上进行的实验。BIAcore 2000仪器将使我们能够直接观察和测量真实的时间的分子相互作用的结合亲和力和缔合/解离动力学，从而大大加快了识别和表征蛋白质-蛋白质和蛋白质-DNA相互作用的过程。BIAcore仪器在传感器芯片上含有固定化配体，并且可以通过芯片表面附近的折射率变化，通过表面等离子体共振检测来监测游离溶液中未标记蛋白质对芯片的吸附。这些变化与吸附质量的变化成正比，从而能够表征生物分子相互作用。结合的材料也可以从传感器芯片洗脱和回收，从而允许识别先前未知的分子相互作用。该仪器还可用于筛选表达scFv序列的噬菌体展示文库，以及表征已知的蛋白质-蛋白质或蛋白质-DNA相互作用。BIAcore 2000将用于支持主要用户正在进行的NIH资助项目，并将大大提高这些研究工作的生产力。

The purposed of this Shared Instrumentation grant proposal is to purchase a Biosensor BIAcore 2000 instrument for a group of seven investigators in the Department of Vascular Biology at The Scripps Research Institute. These investigators are studying protease and protease inhibitor structure and function in the vascular system, integrin structure and function, apolipoprotein structure and function, pro-hormone processing, and gene regulation. The areas of investigation of this group of investigators of this group of investigators are relevant to both physiological and pathological processes which include thrombolysis, atherosclerosis, myocardial infarction, obesity, cancer, hypertension, neuronal function, development and wound healing. These studies emphasize protein-protein and protein-DNA interactions. Since several of these investigations are studying either the same or related molecules, we will able to draw on the experience of other members of the users group in designing experiments to be performed on the BIAcore. The BIAcore 2000 instrument will allow us to directly observe and measure the binding affinities and association/dissociation kinetics of molecular interactions in real time, thus greatly accelerating the process of identifying and characterizing both protein-protein and protein-DNA interactions. The BIAcore instrument contains an immobilized ligand on a sensor chip and can monitor the adsorption of unlabeled proteins in free solution to the chip by means of surface plasmon resonance detection by changes in the refractive index in the vicinity of the surface of the chip. These changes are directly proportional to the change in adsorbed mass, enabling characterization of biomolecular interactions. The bound material can also be eluted and recovered from the sensor chip, thus allowing identification of previously unknown molecular interactions. This instrument can also be used to screen phage display libraries which express scFv sequences, as well as to characterize known protein-protein or protein-DNA interactions. The BIAcore 2000 will be used to support ongoing NIH-funded projects of the major users, and will substantially enhance the productivity of these research efforts.