Purchase of Applied Biosystems QTRAP 5500 Mass Spectrometer

描述（由申请人提供）：本提案寻求购买第三代AB/Sciex QTRAP 5500质谱仪的资金。这台共享仪器将用于对特定的后修饰肽进行可靠的定量，精确到阿托摩尔水平-这是QTRAP 5500明确设计的任务，并且是唯一适合的任务。整体的主要重点是研究特定位点的动力学和蛋白质功能的调节，以响应这一不同的生物医学调查组的各种细胞线索。将为每个主要项目选择共价修饰肽组，通过多反应监测（MRM）进行定量测量。共价添加调节和对接部分（赖氨酰e-氨基功能的磷酸化、O-GlcNAc酰化和甲基化（单-、二-和三-）、乙酰化和泛素化等）是酶促可逆的，但蛋白水解酶的作用产生特定蛋白质组本身的不可逆裂解。该仪器将被安置在UCSF新的使命湾校区的NIH NCRR支持的质谱资源中心。这种环境确保了科学、技术和管理专门知识的可用性和参与，以优化其绩效和研究生产力。主要用户的主要主题集中在研究的动力学后调节的蛋白质，形成和调制的信号通路和网络，以及招聘的蛋白质复合物，如那些调节染色质生物学。研究人员主要需要测量个别网站特定的化学计量和他们的时间调制的细胞线索。他们是一个由六名主要研究人员组成的小组，参与了大约21个NIH支持的研究项目。这些研究项目处于人类生物学及其疾病状态研究的最前沿。Charles Craik博士将担任咨询委员会主席，负责对共用文书的使用提供总体指导。大卫Maltby先生将监督性能优化，调度，培训，维护，并将提供日常监督的使用。与UCSF-NCRR资源的密切联系和主要用户的财政支持将确保长期有效的使用。 公共卫生相关性：蛋白质的化学修饰调节其功能。这些修饰的调节受损在诸如HIV、癌症、阿尔茨海默病和癫痫等疾病中起作用。我们的研究目标是可重复地量化蛋白质修饰的水平，以了解上述疾病的分子原因。

DESCRIPTION (provided by applicant): This proposal seeks funding for the purchase of a third generation AB/Sciex QTRAP 5500 mass spectrometer. This shared instrument will be used for reliable quantitation of specific posttranslationally modified peptides down to the attomole level - a task for which the QTRAP 5500 was explicitly designed and is uniquely suited. The overall major focus is on studies of the site-specific dynamics and modulation of protein function in response to a variety of cellular cues for this diverse group of biomedical investigations. Sets of covalently modified peptides will be selected for each major project for quantitative measurements by multiple reaction monitoring (MRM). Covalent addition of regulatory and docking moieties (phosphorylation, O-GlcNAcylation, and methylation (mono-, di- and tri-), acetylation and ubiquitination of lysyl e-amino functions, etc.) are enzymatically reversible, but the action of proteolytic enzymes produce irreversible cleavages of particular sets of proteins themselves. The instrument will be housed in the NIH NCRR-supported Mass spectrometry Resource Center at the new Mission Bay Campus at UCSF. This environment assures the availability and involvement of the scientific, technical and management expertise required to optimize its performance and research productivity. The primary theme of the major users focuses on studies of the dynamics of posttranslationally regulated proteins that underlies formation and modulation of signaling pathways and networks as well as the recruitment of protein complexes such as those that regulate chromatin biology. Investigators have major needs in measurements of individual site-specific stoichiometries and their temporal modulation by cellular cues. They are a group of six Principal Investigators involved in some twenty-one NIH supported research programs. These research projects are at the forefront of research in human biology and its disease states. Dr. Charles Craik will chair the Advisory Committee responsible for the overall guidance of the shared instrument's use. Mr. David Maltby will oversee optimization of performance, scheduling, training, maintenance and will provide daily supervision of usage. The close association with the UCSF-NCRR resource and the financial support of the major users will assure long-term productive usage. PUBLIC HEALTH RELEVANCE: Chemical modifications of proteins regulate their function. Impaired regulation of these modifications plays a role in such diseases as HIV, cancer, Alzheimer's disease and epilepsy. Our research goal is to reproducibly quantify levels of protein modification to gain an understanding of the molecular causes of the previously mentioned diseases.