Purchase of a MALDI-TOF-TOF Mass Spectrometer

描述（由申请人提供）：申请MALDI-TOF-TOF质谱仪及其相关的液相色谱和定点机器人的资金支持，这对许多nih资助的研究项目至关重要。为了确保最大限度的可用性，适当的维护和NIH资金的最佳使用，该仪器将放置在UCI质谱分析设备中，该设备的主任拥有超过30年的经验。该设施没有MALDI-TOF-TOF。UCI长期以来一直支持该设施，今年的资金大幅增加，增加了一名蛋白质质谱仪的资金，并斥资15万美元对3500平方英尺的空间进行了翻新，以重新安置该设施，包括用于蛋白质样品制备和蛋白质ms的专用房间。此外，仪器预算也投入了9万美元。正常业务资金通过充值获得。该设施设备齐全，使用频繁，特别是使用开放获取技术进行小分子分析，每年通过流动注射ESI， C18-LC-ESI-MS和GC-MS分析20,000个样品。然而，下面的研究表明，在旧的、不再生产的MALDI-TOF（单分析仪）仪器和灵敏度不合适或不够高的QTOF之外，迫切需要扩大蛋白质分析能力。支持的研究项目分布在物理科学学院、生物科学学院和医学院，因此，该仪器将真正成为一个共享的跨学科资源。主要用户的研究项目有：a) Weiss教授（PI）：噬菌体溶解蛋白工程后选择的膜蛋白小窝蛋白突变体测序；用潜在的HIV抑制治疗剂修饰的肽表位的分析，旨在抵消HIV的易变性问题。b) Aswad教授：l -异天冬氨酸甲基转移酶在受损蛋白甲基化和非典型异天冬氨酸残基通过异肽键转化为正常肽键修复中的活性；研究蛋白精氨酸甲基化在基因表达调控中的作用，特别是与精氨酸甲基转移酶4活性有关。c) James教授：蚊子蛋壳蛋白测序，研究伊蚊和按蚊的进化差异，并可能为病媒控制提供新途径。d) Sandri-Goldin教授：理解精氨酸甲基化作为单纯疱疹病毒调节蛋白ICP27的分子开关机制，ICP27是一种多功能蛋白，其活性在感染的时间过程中被修饰。e) Shea教授：表位识别聚合物的发展，它可以像合成抗体一样起作用。f) Walsh教授：凋亡信号转导在免疫系统的发育、激活和稳态中所起的作用。在可能的情况下，其他由nih资助的研究小组也可以使用该仪器。与公共卫生有关：要求为MALDI-TOF-TOF质谱仪和相关设备提供资金，这些设备对于开展分析化学工作以支持各种疾病状态下生物体代谢的基础研究至关重要。这些包括：i)病毒研究，病毒代谢和控制机制，包括艾滋病毒和单纯疱疹；ii)细胞调节，控制细胞活化、细胞死亡和老化蛋白的恢复；iii)昆虫代谢，蚊子进化和疟疾蚊子控制的替代方法；iv)合成抗体的开发。

DESCRIPTION (provided by applicant): Funding support is requested for a MALDI-TOF-TOF mass spectrometer with associated liquid chromatography and spotting robot that are essential to many NIH-funded research programs. To ensure maximum availability, proper maintenance and best use of NIH funds, the instrument will be placed in the UCI Mass Spectrometry Facility, the Director of which has over 30 years experience. The facility does not have a MALDI-TOF-TOF. UCI has long supported the Facility and this has been dramatically increased this year with the addition of funding for a protein mass spectrometrist and a $150,000 renovation of 3,500 square feet of space to re-house the facility that includes dedicated rooms for protein sample preparation and protein MS. Also $90,000 is committed to the instrument budget. Normal operational funds are obtained though recharge. The Facility is well equipped and heavily used, particularly for small molecule analysis using Open Access technology, analyzing 20,000 samples a year by flow injection ESI, C18-LC-ESI-MS and GC-MS. However, the research below shows that there is an urgent need to expand the protein analysis capability beyond the old and no longer manufactured MALDI-TOF (single analyzer) instrument and a QTOF that is not appropriate or sufficiently sensitivity. The research projects to be supported are spread across the Schools of Physical Sciences, Biological Sciences and Medicine and the instrument will, therefore, be truly a shared inter-disciplinary resource. Research projects of the major users include: a) Professor Weiss (PI): Sequencing of mutants of the membrane protein caveolin selected after engineering of phage solubilized protein; analysis of peptide epitopes modified with potentially therapeutic agents for HIV inhibition and designed to counteract the issue of HIV mutability. b) Professor Aswad: Activity of L-isoaspartyl methyltransferase in the methylation of damaged proteins and repair of atypical isoaspartyl residues by conversion of the isopeptide bond to a normal peptide bond; investigating the role of protein-arginine methylation in the regulation of gene expression, particularly concerning arginine methyltransferase 4 activity. c) Professor James: Mosquito egg shell proteins sequencing to study evolutionary divergence of Aedes and Anopheles mosquitoes and potentially a new pathway for vector control. d) Professor Sandri-Goldin: Understanding arginine methylation as a molecular switching mechanism in the herpes simplex virus regulatory protein, ICP27, a multifunctional protein whose activities are modified during the time course of infection. e) Professor Shea: Development of epitope recognition polymers that can act like synthetic antibodies. f) Professor Walsh: The role that apoptotic signal transduction plays in the development, activation and homeostasis of the immune system. Other NIH-funded research groups will be provided access to the instrument when possible. PUBLIC HEALTH RELEVANCE: Funds are requested for a MALDI-TOF-TOF mass spectrometer and associated equipment that is essential for undertaking analytical chemistry in support of fundamental research into organism metabolism in various disease states. These include: i) Virus research, mechanisms of virus metabolism and control including HIV and Herpes simplex; ii) Cell regulation, control of cell activation, cell death and restoration of aging proteins; iii) Insect metabolism, mosquito evolution and alternative methods of malarial mosquito control and iv) Development of synthetic antibodies.