A SHARED BIACORE BIOSENSOR

我们寻求一种共享的Biacore 3000表面等离子体共振生物传感器，可用于三个即时应用：1-展示分子之间的直接物理相互作用，并区分不同候选分子之间物理相互作用的相对强度2-绘制决定与结合伙伴相互作用的氨基酸和接触区域，并量化修饰蛋白质或多肽与其结合伙伴的相对亲和力。为了测试识别复杂混合物中相互作用的分子的可行性，了解决定细胞行为的分子网络需要经过几个步骤。首先，确定影响过程或途径功能的所有元素；其次，确定哪些元素在物理上相互作用；第三，确定一个分子与其他潜在结合伙伴之间相互作用的相对强度；最后，确定改变的条件或环境(例如，pH、浓度、磷酸化等)如何影响分子相互作用的性质。分子相互作用的测量在许多研究领域中越来越重要。对我们小组来说，特别重要的是需要通过直接测量物理相互作用来测试遗传相互作用数据。虽然有几种方法可以用来监测物理相互作用(例如酵母双杂交、免疫共沉淀等)，但这些方法在技术上都具有挑战性。与其他方法相比，生物传感器技术具有快速、灵敏、定量和技术上容易得多的特点。它也是普遍适用的，因为它不依赖于相互作用分子的特定性质，而不是依赖于它们的物理相互作用。此外，一旦使用生物传感器技术检测到相互作用，就可以使用相同的固定化受体和配体制剂来定量测量动力学速率常数和平衡结合常数，从而节省了大量的时间和精力。这里列出的项目(以及其他项目)将从分子相互作用的快速测量中获得巨大好处。

We seek a shared BIAcore 3000 surface plasmon resonance biosensor, to be used for 3 immediate applications; 1 - To demonstrate direct physical interaction between molecules and to distinguish the relative strength of physical interactions between different candidates 2- To map the amino acids and areas of contact that dictate interaction with binding partners and quantitate the relative affinities of modified proteins or peptides with their binding partners. 3- To test the feasibility of identifying interacting molecules in complex mixtures Understanding the molecular networks that dictate cell behavior proceeds through several steps. First, identifying all the elements that affect the function of a process or pathway; second, identifying which elements interact physically; third, identifying the relative strength of the interaction between one molecule and other potential binding partners; and finally, determining how altered conditions or environments (e.g. pH, concentration, phosphorylation etc) affect the nature of molecular interactions. Measurements of molecular interaction are increasingly important in many areas of investigation. Of particular importance to our group is the need to test genetic interaction data by direct measurements of physical interaction. Although several approaches can be used to monitor physical interaction (e.g. yeast two-hybrid, co-immunoprecipitation etc), those methods are all technically challenging. Biosensor technology is fast, sensitive, quantitative and technically much easier than these other approaches. It is also generally applicable in that it does not rely on specific properties of the interacting molecules other than their physical interaction. In addition, once interactions are detected using biosensor technology, quantitative measurements of kinetic rate constants and equilibrium association constants can be made using the same immobilized receptor and ligand preparations, thus saving considerable time and effort. The projects listed here (as well as others) would reap huge benefit from rapid measurements of molecular interactions.