ACQUISITION OF A BIACORE 2000 BIOSENSOR

这项提案来自俄勒冈州州立大学三所学院的五名研究人员，他们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，每种都能够对蛋白质-蛋白质和蛋白质-核酸缔合和解离进行真实的定量分析。 我们选择了BIAcore 2000，这是一种全自动仪器，可以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。 一旦缔合反应完成，然后除去溶液中的蛋白质，并随后解离复合物，其中缔合和解离都在真实的时间内进行。 因此，人们可以确定结合和关闭速率，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。 简单的修改允许分析组装的多蛋白复合物和小分子的结合平衡的影响。 计划的应用包括研究DNA前体生物合成中的蛋白质-蛋白质相互作用（马修斯）、结合质谱分析蛋白质-核酸相互作用（巴罗夫斯基）、植物病毒RNA复制中的RNA-蛋白质相互作用（德雷尔）、配体诱导的维甲酸受体的构象变化（Leid）和纤维素分解酶的亲和配体（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).