A HYBRID 12T Q-FTMS FOR THE INVESTIGATION OF NUCLEIC ACIDS: BIOCHEMISTRY

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：本提案旨在为购置配备12特斯拉超导磁体、电喷雾电离（ESI）、电子捕获解离（ECD）和红外多光子解离（IRMPD）的混合四极/阱-傅立叶变换离子回旋共振质谱仪寻求资金。这种配置提供了完成核酸及其非共价和共价加合物的高分辨率表征所需的最先进的能力，这是所支持的研究项目的核心。参与者建立了旨在通过解决有关逆转录病毒复制和包装，耐药性以及基因损伤和修复的基本问题来促进我们对传染病和癌症的理解的计划。拟议的仪器将提供结构和功能的调查，这将是不可能的结构生物学和生物化学中使用的传统方法的基础。特别是，它将能够实现结构探测、结合常数测定、动力学分析和对降解非常敏感的生物底物（例如，RNA和RNA加合物），固有不稳定（例如，非共价复合物），并且以非常低的量存在（例如，DNA-加合物）。由高磁场提供的分辨能力将是至关重要的复杂的分析物混合物的分析，通过处理与足迹试剂，双功能交联剂，生化探针，致癌剂，核苷酸类似物，和候选抑制剂配体的核酸底物和蛋白质-核酸组件。混合的前端将能够控制允许进入池的离子的数量，以提高串联质谱法的分辨率、动态范围和性能。ECD和IRMPD的组合将提供完成化学修饰产物和交联杂缀合物的明确表征的能力。除了所描述的由主要用户领导的项目外，拟议的仪器将支持许多其他生物医学研究人员的工作，这些研究人员已被证明需要高分辨率质谱仪（次要用户）。预期的服务构成部分将补充两个不同的核心设施的活动，这两个设施分别为马里兰州巴尔的摩县大学校园和巴尔的摩的马里兰州大学格林鲍姆癌症中心和医学院提供服务。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation 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 grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): This proposal seeks funding for the acquisition of a hybrid quadrupole/trap-Fourier transform ion cyclotron esonance mass spectrometer equipped with a 12 Tesla superconductive magnet, electrospray ionization (ESI), electron capture dissociation (ECD), and infrared multiphoton dissociation (IRMPD). This configuration affords the state-of-the-art capabilities necessary to complete the high-resolution characterization of nucleic acids and their non-covalent and covalent adducts, which is at the core of the supported research projects. The participants have established programs aimed at advancing our understanding of infectious diseases and cancer by tackling fundamental questions concerning retroviral replication and packaging, drug resistance, and gene damage and repair. The proposed instrument will provide the basis for structural and functional investigations that would not be possible by traditional approaches used in structural biology and biochemistry. In particular, it will enable the implementation of structural probing, binding constants determination, kinetics analysis, and top-down characterization of biological substrates that are very susceptible to degradation (e.g., RNA and RNA-adducts), inherently labile (e.g., non-covalent complexes), and present in very low amounts (e.g., DNA-adducts). The resolving power afforded by the high magnetic field will be crucial for the analysis of the complex analyte mixtures obtained by treating nucleic acid substrates and protein-nucleic acids assemblies with footprinting reagents, bifunctional crosslinkers, biochemical probes, carcinogenic agents, nucleotide analogues, and candidate inhibitor ligands. The front- end of the hybrid will enable to control the number of ions admitted into the cell to improve resolution, dynamic range, and performance in tandem mass spectrometry. The combination of ECD and IRMPD will provide the ability to complete the unambiguous characterization of chemically-modified products and crosslinked heteroconjugates. In addition to the described projects lead by the primary users, the proposed instrument will support the work of many other biomedical investigators with a proven need for high- resolution mass spectrometry (secondary users). The anticipated service component will complement the activities of two distinct core facilities serving the University of Maryland Baltimore County (UMBC) campus and the University of Maryland at Baltimore (UMB) Greenebaum Cancer Center and Medical School.