Purchase of a MALDI-TOF-TOF Mass Spectrometer

购买 MALDI-TOF-TOF 质谱仪

• 批准号: 7595706
• 负责人: Gregory A. Weiss
• 金额: $ 50万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-14 至 2011-05-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595706
• 关键词: Aedes; Aging; Amino Acid Sequence; Analytical Chemistry; Anopheles Genus; Antibodies; Apoptotic; Arginine; Bacteriophages; Biological Sciences; Budgets; Cell Death; Cells; Commit; Culicidae; Development; Disease; Egg Shell; Engineering; Ensure; Epitopes; Equipment; Evolution; Funding; Funding Applicant; Gene Expression Regulation; HIV; Homeostasis; Housing; Human Herpesvirus 2; Immune system; Infection; Injection of therapeutic agent; Insecta; Liquid Chromatography; Maintenance; Mass Spectrum Analysis; Medicine; Membrane Proteins; Metabolism; Methods; Methylation; Methyltransferase; Molecular; Mosquito Control; Organism; Pathway interactions; Peptide Sequence Determination; Peptides; Play; Polymers; Preparation; Protein Analysis; Proteins; Regulation; Research; Research Project Grants; Resources; Robot; Role; Sampling; Schools; Signal Transduction; Simplexvirus; Spottings; Technology; Therapeutic Agents; Time; United States National Institutes of Health; Virus; arginine methyltransferase; design; experience; fundamental research; genetic regulatory protein; instrument; mass spectrometer; mutant; physical science; professor; programs; public health relevance; repaired; restoration; small molecule; square foot; vector control

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.

描述（由申请人提供）：请求为 MALDI-TOF-TOF 质谱仪提供资金支持，并配备相关的液相色谱仪和点样机器人，这对于许多 NIH 资助的研究项目至关重要。为了确保 NIH 资金的最大可用性、适当维护和最佳利用，该仪器将放置在 UCI 质谱设施中，该设施的主任拥有 30 多年的经验。该设施没有 MALDI-TOF-TOF。 UCI 长期以来一直支持该设施，今年增加了对蛋白质质谱仪的资助，并斥资 150,000 美元翻新了 3,500 平方英尺的空间来重新安置该设施，其中包括用于蛋白质样品制备和蛋白质的专用房间。多发性硬化症。此外，还有 90,000 美元用于仪器预算。正常运营资金通过充值获得。该设施设备齐全，使用频繁，特别是使用开放存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，迫切需要将蛋白质分析能力扩展到旧的和不再制造的 MALDI-TOF（单一分析仪）仪器和不合适或灵敏度不够的 QTOF 之外。所支持的研究项目遍布物理科学学院、生物科学学院和医学院，因此该仪器将成为真正的跨学科共享资源。主要用户的研究项目包括： a) Weiss教授（PI）：对噬菌体溶解蛋白工程后选择的膜蛋白caveolin突变体进行测序；分析用潜在的 HIV 抑制治疗剂修饰的肽表位，旨在解决 HIV 突变问题。 b) Aswad 教授：L-异天冬氨酰甲基转移酶在受损蛋白质甲基化中的活性以及通过将异肽键转化为正常肽键来修复非典型异天冬氨酰残基；研究蛋白质-精氨酸甲基化在基因表达调节中的作用，特别是精氨酸甲基转移酶 4 活性。 c) James 教授：通过蚊子卵壳蛋白测序来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：了解精氨酸甲基化作为单纯疱疹病毒调节蛋白 ICP27 中的分子转换机制，ICP27 是一种多功能蛋白，其活性在感染过程中发生改变。 e) Shea 教授：开发可以像合成抗体一样发挥作用的表位识别聚合物。 f) Walsh 教授：细胞凋亡信号转导在免疫系统的发育、激活和稳态中发挥的作用。如果可能的话，美国国立卫生研究院资助的其他研究小组将能够使用该仪器。公共健康相关性：需要资金购买 MALDI-TOF-TOF 质谱仪和相关设备，这对于进行分析化学以支持各种疾病状态下生物体代谢的基础研究至关重要。其中包括： 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、细胞活化的控制、细胞死亡和老化蛋白质的恢复； iii) 昆虫新陈代谢、蚊子进化和控制疟疾蚊子的替代方法以及 iv) 合成抗体的开发。

项目成果

Hydrazide reactive peptide tags for site-specific protein labeling.

用于位点特异性蛋白质标记的酰肼反应肽标签。

• DOI: 10.1021/bc200415v
• 发表时间: 2011-10-19
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Eldridge, Glenn M.;Weiss, Gregory A.
• 通讯作者: Weiss, Gregory A.