Hybrid Quadrupole-Time of Flight Mass Spectrometer

这是路易斯维尔大学生物分子质谱实验室（UofL）购买和操作混合四极杆飞行时间质谱仪的建议。目前在UofL或该地区还没有这种高分辨率、准确的质谱分析方法。本提案的具体目的如下：提供(1)高分辨率，准确的串联质谱分析能力，用于生物学相关分子的定性鉴定。(2)能够对(2)修饰的DNA和RNA， (b)蛋白质的翻译后修饰和(c)作为生物控制机制或外源毒性标记的非共价蛋白质-蛋白质或蛋白质-核酸相互作用进行高灵敏度结构分析。(3)为在分子水平上研究生物防治机制的定量技术的发展提供了可能。（四）实施科研发展和科学事业发展教育计划。已经确定了四个主要用户研究焦点小组，其中包括由NIH 8个不同研究所资助的16个主要用户。具体的应用是多种多样的，但主要集中在四个领域之一，其中混合Q-TOF技术已经成功整合：(1)细胞-细胞通讯：信号传导和翻译后修饰，(2)病理和毒理学中的氧化过程，(3)功能基因组学和蛋白质组学，(4)分子内分泌学。这些研究科学家将立即受益于Q-TOF的可用性，这些项目已经被NIH审查认为具有科学重要性，并且将有灵活的选择供用户访问。一项教育推广战略将招募其他NIH资助的用户来整合这项技术。路易斯维尔大学坚定地承诺支持这一申请，其总价值是所要求资金的两倍。详细描述了有效管理和使用仪器的机制。大学还承诺支付设置、安装、维护和支持人员的费用。该大学的承诺为期五年。首席研究员将依靠技术咨询小组和内部咨询委员会的意见，其组成在提案中有所规定。该项目将允许这些调查人员和其他人员突破技术障碍，通过内部分析能力满足他们的需求。

This is a proposal to purchase and operate a hybrid quadrupole-time of flight mass spectrometer in the Biomolecular Mass Spectrometry Laboratory of the University of Louisville (UofL). No such high resolution, accurate mass spectrometry for is available now at the UofL or in the region. The Specific Aims of this Proposal are as follows: provision of (1) High resolution, accurate tandem mass spectrometry capabilities for qualitative identification of biologically relevant molecules. (2) Enabling of high sensitivity structural analysis of (2) modified DNA and RNA, (b) post-translational modifications of proteins and (c) non-covalent protein-protein or protein-nucleic acid interactions as biological control mechanisms or markers of xenobiotic toxicity. (3) Enabling of development of quantitative techniques for study of biological control mechanisms at the molecular level. (4) Implementation of an education program for research development and scientific career development. Four Major Users Research Focus Groups have been identified, including 16 Major Users funded by 8 different Institutes of the NIH. The specific applications are quite varied, but are focused in one of four areas in which hybrid Q-TOF technology has been successfully integrated: (1) Cell-cell communications: Signaling and Post- translational modifications, (2) Oxidative processes in pathology and toxicology, (3) Functional Genomics and Proteomics and (4) Molecular Endocrinology. These research scientists will benefit immediately from availability of the Q-TOF for the projects already deemed scientifically important by NIH review, and will have flexible options for user access. An educational outreach strategy will recruit other NIH funded users to incorporate this technology. The University of Louisville makes a strong commitment in support of this application, with an overall value of twice the funds requested. A detailed mechanism is described for efficient management and use of the instrument. The University also commits to paying for the costs of setup, installation, maintenance and support personnel. The commitment of the University is made for a period of five years. The Principal Investigator will rely on advice from a Technical Advisory Group and an Internal Advisory Committee, the composition of which is specified in the proposal. This project will allow these investigators and others to break through a technical roadblock to have their requirements met by in-house analytical capability.