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

描述(申请人提供)：此申请是为了获得最先进的Nano-LC-MS/MS离子陷阱质谱仪，这是Bruker Daltonics的Amazon ETD，将为蛋白质组学翻译后修饰提供准确、可重复性和高灵敏度的测量。该仪器将为我们的质谱学核心带来显著的增强，并将支持多个NIH资助用户的研究努力。除了高度的灵敏度外，电子转移解离(ETD)作为一种碎裂技术的加入将促进蛋白质组学在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)方面的新发现，例如慢性氧化应激导致的蛋白质加合物以及由此产生的脂质过氧化产物4-羟基壬烯醛和丙烯醛。美国国立卫生研究院资助的多名科罗拉多大学丹佛分校的研究人员被提议共享该仪器的使用。获得这一仪器的主要理由是加强这些由美国国立卫生研究院资助的蛋白质组学发现领域的研究项目，利用增加的敏感性和ETD碎片作为关键组件。该应用程序的调查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，该仪器拟设在药学院，该学院致力于支持和开发其质谱学核心，并将为仪器维护和维修提供必要的服务合同，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。正如本申请中详细描述的那样，每个主要用户都展示了NIH资助的研究的丰富和多产的历史。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本生理方面所起的作用。用碰撞诱导解离(CID)和ETD碎裂技术检测这些修饰的能力将增加修饰多肽的覆盖率。特别是，在标准CID裂解条件下不稳定的蛋白质修饰将通过使用ETD来表征更多的细节。这些改进，再加上我们现有离子陷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：这一应用的仪器是来自Bruker Daltonics的Amazon ETD/CID离子陷阱质谱仪和纳米液相色谱系统，它将为蛋白质组学翻译后修饰提供准确、可重复性和高灵敏度的测量。该仪器将支持美国国立卫生研究院资助的多项研究工作，在科罗拉多大学丹佛健康科学校区表征疾病状态改变的蛋白质组学。 购买这一仪器将大大加强共享用户在研究神经退行性疾病、肝脏疾病和癌症的病理学方面的短期和长期研究目标。

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.