A sensitive, ETD capable, ion trap for proteomics and PTM research

用于蛋白质组学和 PTM 研究的灵敏、具有 ETD 功能的离子阱

• 批准号: 8247444
• 负责人: DENNIS PETERSEN
• 金额: $ 38.78万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-05 至 2013-06-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247444
• 关键词: 4 hydroxynonenal; Acrolein; Area; Chronic; Colorado; Contract Services; Core Facility; Coupled; Disease; Dissociation; Electron Transport; Evaluation; Fees; Funding; Goals; Housing; Ions; Link; Lipid Peroxidation; Liver diseases; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Modification; Neurodegenerative Disorders; Oxidative Stress; Pathology; Peptides; Pharmacy Schools; Phosphorylation; Physiology; Play; Post-Translational Protein Processing; Protein Acetylation; Proteins; Proteomics; Recording of previous events; Recovery; Research; Research Personnel; Research Project Grants; Research Support; Techniques; United States National Institutes of Health; Universities; adduct; cost; experience; instrument; instrumentation; mass spectrometer; nano; novel; repaired; skills

DESCRIPTION (provided by applicant): This application is for acquisition of a state-of-the-art nano-LC-MS/MS ion trap mass spectrometer, an amaZon ETD from Bruker Daltonics, which will provide accurate, reproducible and highly sensitive measurement of proteomics post-translational modifications. This instrument will bring significant enhancements to our Mass Spectrometry Core and will support the research efforts of multiple NIH-funded users. In addition to the high levels of sensitivity, the addition of electron transfer dissociation (ETD) asa fragmentation technique will facilitate novel proteomics discoveries in protein phosphorylation and acetylation as well as numerous other significant post-translational modifications (PTMs) such as protein adducts due to chronic oxidative stress and the resulting lipid peroxidation products 4-hydroxynonenal and acrolein. Multiple NIH-funded researchers at the University of Colorado Denver are proposed to share the use of this instrument. The primary justification for acquiring this instrument is to enhance these NIH-funded research projects in the area of proteomics discovery, utilizing the added sensitivity and ETD fragmentation as critical components. The investigators in this application possess the required experience and technological skill-set to maintain and properly use this instrument. Furthermore, the School of Pharmacy, where this instrument is proposed to be housed, is dedicated to supporting and developing its Mass Spectrometry Core and will provide the requisite service contracts for instrument maintenance and repair, as it does for each instrument housed within the Core facility. Furthermore, a cost recovery fee plan guarantees the financial viability for the instrumet over the long term. As detailed within this application, each major user demonstrates a rich and productive history in NIH-funded research. These NIH-funded projects range from neurodegenerative diseases to cancer and liver disease. A common denominator linking each of these projects is the need for highly sophisticated proteomics evaluation of PTMs and the impact these modifications play in altering basal physiology. The ability to examine these modifications with both collision induced dissociation (CID) and ETD fragmentation techniques will provide enhanced coverage of modified peptides. In particular, protein modifications which are labile under standard CID fragmentation conditions will be characterized with increased detail using ETD. These enhancements, coupled with an increase in sensitivity by 50-fold over our existing ion trap instrumentation will provide the capabilities for each shared user to reach both short and long-term research goals of studying altered proteomics under a wide variety of disease pathologies. PUBLIC HEALTH RELEVANCE: The instrument for this application is an amaZon ETD/CID ion trap mass spectrometer and nano liquid chromatography system from Bruker Daltonics which will provide accurate, reproducible and highly sensitive measurement of proteomics post-translational modifications. This instrument will support multiple NIH-funded research efforts in characterizing disease state-altered proteomics at the University of Colorado Denver Health Sciences Campus. Acquisition of this instrument would significantly enhance the efforts of the shared users in meeting both short and long-term research objectives in studying pathologies of neurodegenerative disease, liver disease and cancer.

描述（由申请人提供）：本申请用于购买最先进的纳米 LC-MS/MS 离子阱质谱仪，这是布鲁克道尔顿 (Bruker Daltonics) 的亚马逊 ETD，它将提供准确、可重复且高度灵敏的测量蛋白质组学翻译后修饰。该仪器将为我们的质谱核心带来显着增强，并将支持多个 NIH 资助的用户的研究工作。除了高水平的灵敏度之外，电子转移解离 (ETD) 作为片段化技术的添加将促进蛋白质磷酸化和乙酰化以及许多其他重要的翻译后修饰 (PTM) 的新蛋白质组学发现，例如由于慢性氧化应激和由此产生的脂质过氧化产物4-羟基壬烯醛和丙烯醛。科罗拉多大学丹佛分校的多名 NIH 资助的研究人员提议共享该仪器的使用。购买该仪器的主要理由是利用增加的灵敏度和 ETD 碎片作为关键组件，增强 NIH 资助的蛋白质组学发现领域的研究项目。本申请的研究人员拥有维护和正确使用该仪器所需的经验和技术技能。此外，拟安置该仪器的药学院致力于支持和开发其质谱核心，并将提供必要的仪器维护和维修服务合同，就像对核心设施内的每台仪器所做的那样。此外，成本回收费用计划保证了该工具的长期财务可行性。正如本申请中详细介绍的，每个主要用户都在 NIH 资助的研究中展示了丰富而富有成果的历史。这些由美国国立卫生研究院资助的项目范围从神经退行性疾病到癌症和肝脏疾病。连接这些项目的一个共同点是需要对 PTM 进行高度复杂的蛋白质组学评估以及这些修饰在改变基础生理学方面的影响。通过碰撞诱导解离 (CID) 和 ETD 片段化技术检查这些修饰的能力将增强修饰肽的覆盖范围。特别是，在标准 CID 片段化条件下不稳定的蛋白质修饰将使用 ETD 进行更详细的表征。这些增强功能加上我们现有离子阱仪器的灵敏度提高了 50 倍，将为每个共享用户提供能力，以实现研究各种疾病病理下改变的蛋白质组学的短期和长期研究目标。 公共健康相关性：此应用的仪器是亚马逊 ETD/CID 离子阱质谱仪和 Bruker Daltonics 的纳米液相色谱系统，它将提供准确、可重复和高度灵敏的蛋白质组翻译后修饰测量。该仪器将支持 NIH 资助的多项研究工作，以描述科罗拉多大学丹佛健康科学校区疾病状态改变的蛋白质组学特征。 购买该仪器将显着增强共享用户在神经退行性疾病、肝病和癌症病理学研究方面实现短期和长期研究目标的努力。