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，旨在扩大分析能力，使用可广泛使用的仪器对多达数百个残基的RNA/DNA寡聚体进行快速，特异性和灵敏度表征。 这项工作旨在推进国家的最先进的寡核苷酸表征质谱，特别强调离子阱和飞行时间质谱分析。 离子/离子化学在这项工作中起着重要的作用，作为离子转化和结构表征的手段。 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用于后续分析的反应的示例。 电子转移反应是可以在一级结构表征中发挥直接作用的反应的示例。 使用多维MS来最大化结构信息，探索了宽范围的母离子类型的束型和离子阱碰撞激活。 离子/离子反应与这些活化方法的结合将为RNA/DNA寡聚体的表征提供异常广泛的可能性。 质谱法目前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，脂质组学和其他大规模测量挑战等领域。 然而，质谱法用于表征核酸的转录后修饰的强度尚未发展到例如它们用于蛋白质的程度。 因此，这项工作对推进核酸研究具有重大潜力。 一组不同的研究生和本科生从事这个项目将接触到一个高度多学科的调查线，涉及离子物理，离子化学，仪器和分析化学，除了分子生物学。

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.