ACQUISITION OF A BIACORE 2000 BIOSENSOR

俄勒冈州立大学（Oregon State University）三所学院的五名研究人员提出了这项建议，将允许购买自动光学生物传感器。市场上有两种相互竞争的生物传感器仪器，每一种都能够实时进行蛋白质-蛋白质和蛋白质-核酸结合和解离的定量分析。我们选择了BIAcore 2000，这是一种全自动仪器，用于测量称为表面等离子体共振的光学特性。实际上，这意味着可以跟踪蛋白质与先前固定的蛋白质或核酸的结合。一旦结合反应完成，人们就会在溶液中去除蛋白质，并跟踪复合物的解离，同时实时跟踪结合和解离。因此，可以确定结合的开断速率、解离常数和结合化学计量。测量使用少量的材料，并且该过程是自动化的，用于重复的无监控分析。简单的修饰允许分析多蛋白复合物的组装和小分子对结合平衡的影响。计划中的应用包括研究DNA前体生物合成中的蛋白质-蛋白质相互作用（Mathews），结合质谱分析蛋白质-核酸相互作用（Barofsky），植物病毒RNA复制中的RNA-蛋白质相互作用（Dreher），类视黄醇受体中配体诱导的构象变化（Leid），以及纤维素水解酶的亲和配体（Penner）。

This proposal, from five investigators in three colleges at Oregon State University, would allow purchase of an automated optical biosensor. Two competing biosensor instruments are on the market, each capable of performing quantitative analysis, in real time, of protein-protein and protein-nucleic acid associations and dissociations. We have chosen the BIAcore 2000, a fully automated instrument that measures an optical property called surface plasmon resonance. In practical terms, this means that one can follow binding of a protein to a previously immobilized protein or nucleic acid. Once an association reaction is complete, one then removes the protein in solution and follows dissociation of the complex, with both association and dissociation followed in real time. Thus, one can determine binding on and off rates, dissociation constants, and binding stoichiometries. The measurements use small amounts of material, and the process is automated, for repetitive unmonitored analyses. Simple modifications permit analysis of assembly of multiprotein complexes and the effects of small molecules on binding equilibria. Planned applications include research on protein-protein interactions in DNA precursor biosynthesis (Mathews), analyses of protein-nucleic acid interactions in conjunction with mass spectrometry (Barofsky), RNA-protein interactions in plant virus RNA replication (Dreher), ligand-induced conformational changes in retinoid receptors (Leid), and affinity ligands for cellulolytic enzymes (Penner).