LTQ-Orbitrap Mass Spectrometer for Yale University Keck Laboratory

描述（由申请人提供）：本申请要求资助一种最先进的共享混合质谱仪，该质谱仪将为耶鲁大学提供合作研究和核心服务使用。凯克生物技术资源实验室的MS和蛋白质组学核心。由Thermo Fisher Scientific制造的LTQ-Orbitrap混合质谱仪将线性四极杆离子阱与轨道阱质谱仪相结合。当这种要求的质谱仪与我们内部开发的磷酸肽富集方法和现有的“惰性”毛细管纳米级HPLC系统一起使用时，所得到的平台和应用的方法将提供我们认为是第一个强大的，专用的磷蛋白分析和相关的磷酸蛋白质组学服务，可从主要的学术核心设施获得。LTQ-Orbitrap质谱平台具有独特的MS 2和MS 3数据依赖性磷酸肽采集能力，与稳健、高灵敏度、快速扫描、高分辨率、高质量准确度和宽动态范围特性紧密结合，共同创造了无与伦比的能力，可以自信地从高度复杂的消化细胞和组织蛋白提取物中检测和鉴定大量磷酸肽。此外，该平台还提供其他蛋白质翻译后修饰、脂质、代谢物的检测和鉴定，以及凯克实验室目前未提供的其他类型的分析。由于这些独特的能力和磷酸化在控制蛋白质功能中的重要作用，人们对为凯克实验室提供这个平台非常感兴趣，因此它可以在服务费的基础上支持33个生物医学研究项目，这些项目由来自13所大学和医疗中心的26个主要用户和5个次要用户监督。这些用户包括四个NIH中心的成员，这些中心与凯克实验室密切相关，并且具有强大的蛋白质组学组件（即，耶鲁/NHLBI蛋白质组学中心，耶鲁/NIDA神经蛋白质组学中心，NCI资助的耶鲁癌症中心的蛋白质组学共享资源，以及NIAID资助的东北生物防御卓越中心的蛋白质组学核心）。两个主要的用户，理查德利夫顿（NHLBI）和安格斯奈恩（NIDA），将与我们的中心合作，以追求他们在肾脏疾病和药物成瘾和抑郁症的研究应用，同时也合作生物技术研究，旨在进一步推进我们非常有前途的磷酸蛋白质组学技术。这项工作将作为快速开发新的磷酸蛋白质组学服务的基础，这些服务将提供给凯克实验室的所有其他研究贡献者和用户。 相关性：如果这笔赠款得到资助，LTQ-Orbitrap系统将为生物医学研究做出独特的贡献，将扩展到研究疾病的基本机制的研究人员，包括精神分裂症，抑郁症，药物滥用，早产，哮喘，黑色素瘤，肾脏疾病以及生物防御相关微生物病原体的影响，如炭疽。

DESCRIPTION (provided by applicant): This application requests funding for a state-of-the-art shared hybrid mass spectrometer that will provide collaborative research and core service usage for the Yale University, W.M. Keck Biotechnology Resource Laboratory's MS & Proteomics Core. The LTQ-Orbitrap hybrid mass spectrometer manufactured by Thermo Fisher Scientific combines a linear quadrupole ion-trap with the orbitrap mass spectrometer. When this requested mass spectrometer is utilized with our in-house developed phosphopeptide enrichment methodologies and an existing, "inert" capillary nano scale HPLC system, the resulting platform and the applied methodologies will provide what we believe to be the first robust, dedicated phosphoprotein profiling and related phosphoproteomics services available from a major academic core facility. The LTQ-Orbitrap mass spectrometry platform has a unique, MS2 and MS3 data dependent phosphopeptide acquisition capability that is tightly coupled with robust, highly sensitive, fast scanning, high resolution, high mass accuracy, and wide dynamic range characteristics that together create an unmatched ability to confidently detect and identify large numbers of phosphopeptides from highly complex, digested cell and tissue protein extracts of interest. In addition, the platform also offers detection and identification of other protein post-translational modifications, lipids, metabolites, and other types of analyses not currently offered by the Keck Laboratory. Because of these unique capabilities and the important role that phosphorylation plays in controlling protein function, there is very strong interest in providing the Keck Laboratory with this platform so it can be made available on a service charge basis to support 33 biomedical research projects overseen by 26 major and five minor users from 13 universities and medical centers. These users include members of four NIH Centers that are closely associated with the Keck Laboratory and that have strong proteomics components (i.e., Yale/NHLBI Proteomics Center, Yale/NIDA Neuroproteomics Center, the Proteomics Shared Resource in the NCI-funded Yale Cancer Center, and the Proteomics Core of the NIAID-funded Northeast Center of Excellence for Biodefense). Two of the major users, Richard Lifton (NHLBI) and Angus Nairn (NIDA), will collaborate with our center to pursue their research applications in renal disease and in drug addiction and depression while also collaborating on biotechnology research directed at further advancing our highly promising phosphoproteomics technologies. This effort will then serve as the basis for rapidly developing new phosphoproteomics services that will be made available to all other research contributors and users of the Keck Laboratory. Relevance: If this grant is funded, the LTQ-Orbitrap system would make a unique contribution to biomedical research that would extend to investigators studying fundamental mechanisms of diseases including schizophrenia, depression, substance abuse, preterm birth, asthma, melanoma, renal diseases, and the effects of biodefense related microbial pathogens such as anthrax.