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）：资金支持要求为MALDI-TOF-TOF质谱仪与相关的液相色谱和点样机器人，是必不可少的许多NIH资助的研究计划。为了确保最大限度地利用、适当维护和最好地利用NIH的资金，该仪器将被放置在UCI质谱设施中，该设施的主任拥有30多年的经验。该机构没有MALDI-TOF-TOF。UCI长期以来一直支持该设施，今年随着蛋白质质谱仪的资金增加以及3，500平方英尺空间的150，000美元翻新，以重新安置该设施，其中包括蛋白质样品制备和蛋白质MS的专用房间。通过充值获得正常运营资金。该设施设备齐全，使用量很大，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。以下研究表明，迫切需要将蛋白质分析能力扩展到旧的且不再生产的MALDI-TOF之外（单个分析仪）仪器和QTOF不合适或灵敏度不够。支持的研究项目分布在物理科学，生物科学和医学学院，因此，该仪器将真正成为共享的跨学科资源。主要用户的研究项目包括：a）韦斯教授（PI）：对噬菌体溶解蛋白工程化后选择的膜蛋白小窝蛋白突变体进行测序;分析用潜在的治疗剂修饰的肽表位，用于抑制艾滋病毒，并设计用于对抗艾滋病毒突变性问题。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的活性;研究蛋白质-精氨酸甲基化在基因表达调节中的作用，特别是关于精氨酸甲基转移酶4活性。詹姆斯教授：蚊子卵壳蛋白测序研究伊蚊和按蚊的进化差异，并可能为媒介控制提供新途径。（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ICP 27的分子转换机制，ICP 27是一种多功能蛋白，其活性在感染过程中发生变化。e）Shea教授：开发可以像合成抗体一样起作用的表位识别聚合物。f）沃尔什教授：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。其他NIH资助的研究小组将在可能的情况下获得该仪器。公共卫生相关性：要求提供资金，以购置一台MALDI-TOF-TOF质谱仪和相关设备，这对于进行分析化学以支持对各种疾病状态下的有机体代谢进行基础研究至关重要。这些措施包括：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病毒; ii）细胞调节，控制细胞活化，细胞死亡和衰老蛋白的恢复; 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。

项目成果

Hydrazide reactive peptide tags for site-specific protein labeling.

• 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.