Top-Down Tandem Mass Spectrometry of 20-200 Residue RNA and DNA Oligomers

普渡大学的Scott McLuckey教授得到了分析和表面化学计划的支持，该计划的研究旨在扩大分析能力，利用可广泛获得的仪器对多达数百个残基的RNA/DNA寡聚体进行快速、特定和灵敏的表征。这项工作旨在推进最先进的寡核苷酸的质谱学表征，特别是离子陷阱和飞行时间质谱分析。离子/离子化学作为离子转化和结构表征的一种手段，在这项工作中起着重要的作用。单质子转移、金属离子转移和电荷反转是可以将分析物离子从一种形式转化为另一种形式以供后续分析的反应的例子。电子转移反应是可以在一级结构表征中起直接作用的反应的例子。使用多维MS探索了束流类型和离子陷阱对多种母离子类型的碰撞激活，以最大限度地获得结构信息。离子/离子反应与这些活化方法的结合将为表征RNA/DNA低聚物提供不同寻常的广泛可能性。目前，质谱学在现代分子生物学研究中发挥着至关重要的作用，特别是在蛋白质组学、代谢组学、脂类组学等大规模测量挑战领域。然而，质谱学用于表征核酸转录后修饰的强度还没有发展到像蛋白质那样的程度。因此，这项工作对推进核酸研究具有巨大的潜力。从事这个项目的不同的研究生和本科生将接触到涉及离子物理、离子化学、仪器和分析化学以及分子生物学的高度多学科的探究。

Prof. Scott McLuckey at Purdue University is supported by the Analytical and Surface Chemistry Program for investigations aimed at expanding analytical capabilities for the rapid, specific, and sensitive characterization of RNA/DNA oligomers with up to several hundred residues using instrumentation that can be widely available. This work seeks to advance the state-of-the-art in oligonucleotide characterization by mass spectrometry with particular emphasis placed on ion trap and time-of-flight mass analysis. Ion/ion chemistry plays a major role in this work as a means for ion transformation and structural characterization. Single proton transfer, metal ion transfer, and charge inversion are examples of reactions that can transform analyte ions from one form to another for subsequent analysis. Electron transfer reactions are examples of reactions that can play a direct role in primary structure characterization. Both beam-type and ion trap collisional activation of a wide range of parent ion types are explored using multidimensional MS to maximize structural information. The combination of ion/ion reactions with these activation methods will provide an unusually wide range of possibilities for the characterization of RNA/DNA oligomers. Mass spectrometry currently plays a critical role in modern molecular biology research, particularly in the areas such as proteomics, metabolomics, lipidomics, and other large-scale measurement challenges. The strengths of mass spectrometry for characterizing post-transcriptional modifications of nucleic acids, however, have not yet been developed to the extent that they have for proteins, for example. This work, therefore, has significant potential for advancing nucleic acids research. A diverse set of graduate and undergraduate students engaged in this project will be exposed to a highly multi-disciplinary line of inquiry involving ion physics, ion chemistry, instrumentation, and analytical chemistry, in addition to molecular biology.