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

描述（由申请人提供）：本申请旨在获得最先进的nano-LC-MS/MS离子阱质谱仪（来自Bruker Daltonics的amaZon ETD），该质谱仪将提供蛋白质组学翻译后修饰的准确、可重现和高灵敏度测量。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资助用户的研究工作。除了高水平的灵敏度外，电子转移解离（ETD）阿萨片段化技术的加入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引起的蛋白质加合物。科罗拉多丹佛大学的多名NIH资助的研究人员被提议共享该仪器的使用。获得该仪器的主要理由是利用增加的灵敏度和ETD片段作为关键组件，增强这些NIH资助的蛋白质组学发现领域的研究项目。本申请中的研究者拥有维护和正确使用该仪器所需的经验和技术技能。此外，拟将该仪器安置在药学院，药学院致力于支持和开发其质谱核心，并将提供仪器维护和修理所需的服务合同，就像它为核心设施内的每台仪器所做的那样。此外，成本回收收费计划保证了仪器的长期财务可行性。正如本应用程序中所详细介绍的那样，每个主要用户都展示了NIH资助研究的丰富和富有成效的历史。这些NIH资助的项目范围从神经退行性疾病到癌症和肝病。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。用碰撞诱导解离（CID）和ETD片段化技术两者检查这些修饰的能力将提供修饰肽的增强的覆盖。特别地，在标准CID片段化条件下不稳定的蛋白质修饰将使用ETD以增加的细节进行表征。这些增强功能，加上灵敏度比我们现有的离子阱仪器提高了50倍，将为每个共享用户提供能力，以达到在各种疾病病理下研究改变的蛋白质组学的短期和长期研究目标。 公共卫生相关性：用于该应用的仪器是来自Bruker Daltonics的amaZon ETD/CID离子阱质谱仪和纳米液相色谱系统，其将提供蛋白质组学翻译后修饰的准确、可再现和高度灵敏的测量。该仪器将在科罗拉多丹佛大学健康科学校区支持多项NIH资助的疾病状态改变蛋白质组学研究工作。 获得这一仪器将大大加强共享用户在研究神经退行性疾病、肝病和癌症病理学方面的短期和长期研究目标方面的努力。

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.