DIRECT MEASUREMENT OF BINDING AFFINITIES FOR DNA-METAL CATION INTERACTIONS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 我们开发了一种新的，基于质谱的方法来测量特定的金属离子结合到DNA，特别是四链体DNA的亲和常数。 该方法可应用于其它系统，其利用来自结合和未结合物质的气相信号作为确定各种参数的数学模型的输入，所述气相信号通过质谱法测量  其中之一是结合亲和力常数  用非线性最小二乘拟合算法 用于开发和测试模型的系统是凝血酶结合适体，一种结合K+和Sr 2+阳离子的适当四链体结构。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We developed a new, mass spectrometry-based method for measuring affinity constants for specific metal ion binding to DNA, particularly to quadruplex DNA. This method, which is applicable to other systems, utilizes the gas-phase signals, as measured by mass spectrometry, from the bound and unbound species as input into a mathematical model that determines various parameters  one of which is the binding affinity constant  with a non-linear least squares fitting algorithm. The system used to develop and test the model was the thrombin-binding aptamer, an appropriate quadruplex structure that binds both K+ and Sr2+ cations.