Acquisition of a LTQ Orbitrap XL ETD Hybrid FT Mass Spectrometer

描述（由申请人提供）：申请资金用于Thermo Scientific的LTQ Orbitrap XL ETDTM混合FT质谱仪，用于国防，疫苗开发的重要研究，以及了解化学毒性，激酶信号传导和疾病的致病性。蛋白质组学的液相色谱分离，特别是与灵敏和非常准确的串联质谱法相结合，以及与自上而下的蛋白质气相测序相结合的全蛋白质精确测量，是亚利桑那大学现有NIH资助的研究人员非常需要的蛋白质组学能力。该仪器将为我们的LC-MS应用增加价值，特别是对于那些绝对需要增加肽测序置信度的应用，因为碎片离子的精确质量测量（约2 ppm），使用ETD鉴定PTM，鉴定更高电荷的交联肽，以及小蛋白的全蛋白质质量测量。该系统将加入亚利桑那大学亚利桑那蛋白质组学联盟现有的仪器基础设施。获得这种仪器将大大提高我们研究的生产力，这对于NIH资助的研究人员支持这一应用至关重要，他们过去都使用当前的仪器进行了LC-MS，但现在需要高通量精确的蛋白质组学质谱仪。目前还没有这样的蛋白质组学系统存在，尽管可以在校园进行各种质谱分析。Thermo Scientific LTQ Orbitrap XL ETDTM混合FT质谱仪已被证明在蛋白质组学应用中非常可靠。该仪器的获得得到了NIH资助的亚利桑那大学化学和生物化学系（科学医学学院（医学院），分子和细胞生物学（理学院），药理学和毒理学（药学院），生理学（医学院）的研究人员以及其他NIH资助的外部用户的支持。亚利桑那州蛋白质组学联盟还将使该仪器可用于校园内的许多未成年人用户，他们需要这里提出的其他此类特定用途。使用这一升级仪器的战略研究重点领域将是（1）混合物中蛋白质或太平洋西南地区生物防御和新兴疾病卓越中心成员的纯化蛋白质的鉴定和表征，（2）交联肽和蛋白质的表征，以及（3）自上而下的小蛋白质研究。制造商的演示数据和我们的几个用户的演示数据清楚地展示了这种质谱仪的需求和实用性。亚利桑那大学的支持系统在经验丰富的人员、组织和管理能力以及使用记录方面存在，使得收购LTQ Orbitrap XL ETDTM混合FT质谱仪成为应对未来MS研究和科学挑战的合理选择。 公共卫生相关性：使用质谱法进行蛋白质鉴定在很大程度上依赖于获得极其准确的质量测量的能力。所要求的仪器将大大提高我们这样做的能力，并以更大的信心识别对国防，疫苗开发重要的蛋白质，以及那些对增加我们对化学毒性，蛋白质信号传导和疾病致病性的理解重要的蛋白质。所要求的质谱仪将适合亚利桑那州蛋白质组学联盟提供的环境蛋白质组学工具和协议，该联盟由8名员工组成，其中包括博士，MS和BS级科学家。此外，所要求的仪器将被用作培训工具，像我们所有的硬件，为当前和未来的本科生和研究生积极从事健康相关的研究。

DESCRIPTION (provided by applicant): Funds are requested for a LTQ Orbitrap XL ETDTM hybrid FT mass spectrometer, from Thermo Scientific to be used in research important in national defense, vaccine development, and for understanding chemical toxicity, kinase signaling, and pathogenicity of disease. Liquid chromatography separations of protein digests, specifically coupled to sensitive and very accurate tandem mass spectrometry, and whole protein accurate measurements coupled to top-down protein gas-phase sequencing, are highly needed proteomics capabilities for existing NIH-funded investigators at the University of Arizona. This instrument will add value to our LC-MS applications, specifically for those that absolutely require increased confidence of peptide sequencing due to accurate mass measurements of fragment ions (~2 ppm), identification of PTMs using ETD, identification of more highly-charged cross-linked peptides, as well as whole protein mass measurements of small proteins. This system will join the existing instrumentation infrastructure of the Arizona Proteomics Consortium at the University of Arizona. Obtaining this instrument will greatly increase the productivity of our research, and it is critical for the NIH-funded investigators supporting this application, who have all performed LC-MS in the past using current instrumentation but who now have a need for a high-throughput accurate proteomics mass spectrometer. Currently no such proteomics system exists, despite the variety of mass spectrometric analyses that can be performed on campus. The Thermo Scientific LTQ Orbitrap XL ETDTM hybrid FT mass spectrometer has been shown to be extremely reliable for proteomics applications. Acquisition of this instrument is being supported by NIH-funded investigators at the University of Arizona from the Department of Chemistry and Biochemistry, (College of Science Medicine (College of Medicine), Molecular and Cellular Biology (College of Science), Pharmacology and Toxicology (College of Pharmacy), Physiology (College of Medicine), as well as by various other NIH-funded external users. The Arizona Proteomics Consortium will also make the instrument available for numerous minor users on campus who have a need for other such specific uses as proposed here. The strategic research areas of focus for use of this upgraded instrument will be (1) identification and characterization of proteins in mixtures or of purified proteins of members of the Pacific Southwest Regional Center for Excellence in Biodefense and Emerging Diseases, (2) characterization of cross-linked peptides and proteins, and (3) top-down investigations of small proteins. Manufacturer demonstration data and that from several of our users are presented which clearly showcase the need and utility for this mass spectrometer. The support system at the University of Arizona exists in terms of experienced personnel, organizational and management abilities and usage track record, making acquisition of the LTQ Orbitrap XL ETDTM hybrid FT mass spectrometer a logical choice to handle the MS-based research and scientific challenges of the future. PUBLIC HEALTH RELEVANCE: Protein identification using mass spectrometry relies heavily on the ability to obtain extremely accurate mass measurements. The requested instrument will greatly enhance our ability to do just that and to identify with greater confidence proteins important for national defense, vaccine development, and those important for increasing our understanding of chemical toxicity, protein signaling, and pathogenicity of disease. The requested mass spectrometer will fit into the fleet of environmentally proteomic tools and protocols available at the Arizona Proteomics Consortium, which is staffed by eight employees consisting of Ph.D, MS and BS-level scientists. In addition, the requested instrument will be used as a training tool, like all the rest of our hardware, for current and future undergraduate and graduate-level students actively engaging in health-related research.