Purchase of a nanoLC-QTRAP Mass Spectrometer for Targeted Proteomics Studies

购买 nanoLC-QTRAP 质谱仪用于靶向蛋白质组学研究

• 批准号: 8447770
• 负责人: Sonja Hess
• 金额: $ 60万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447770
• 关键词: Affect; Antibodies; Biological; Cell physiology; Complex; Coupled; Cullin Proteins; Embryo; Enzymes; Functional disorder; Funding; Gene Expression; Gene Proteins; Health; Histones; Human; Immunoblotting; Laboratories; Logic; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Mitochondria; Molecular; Monitor; Mus; Mutate; Mutation; Nerve Degeneration; Neurologic; Orthologous Gene; Pathogenesis; Pathway interactions; Peptides; Proteins; Proteome; Proteomics; Research Personnel; Research Project Grants; Risk; Saccharomycetales; Sampling; Schizophrenia; Science; Sea Urchins; Secure; Synapses; System; T cell differentiation; Therapeutic; Woman; Yeasts; base; chemical genetics; genetic regulatory protein; human disease; immune activation; insight; instrument; mass spectrometer; mouse model; multiple reaction monitoring; offspring; response; transcription factor; ubiquitin ligase

DESCRIPTION (provided by applicant): This application is to secure funds to purchase an AB SCIEX QTRAP 5500 PRO triple quadrupole (QQQ) mass spectrometer coupled with an Eksigent nanoLC to carry out analyses based on multiple- reaction monitoring (MRM). MRM is an extremely sensitive, quantitative, and reproducible method for monitoring the levels of targeted, low abundance proteins in a complex biological sample. Conceptually it is similar to performing quantitative immunoblots but without the need to tag or raise antibodies against the targeted proteins. In the long term, this instrument will enable many research projects for which quantitative protein science is a core component. This application comprises eight specific projects. The Deshaies laboratory will quantify the levels of 64 proteins from the cullin-RING ubiquitin ligase (CRL) network in yeast bearing mutations in the Nedd8/Rub1 conjugating and deconjugating enzymes. This analysis will provide insights into the molecular logic of this key regulatory system which has been implicated in the pathogenesis of cancer. The Rothenberg laboratory will study a defined set of transcription factors in mice that will provide fundamental insight in T-cell differentiation. In addition, the Davidson laboratory will study a defined set of transcription factors in sea urchins that will delineate the spatial specifications of gene expression in sea urchin embryos. The Kennedy laboratory will use MRM to quantify synaptic regulatory proteins. The Hess lab will use mass spectrometry to quantify select acetylated histone peptides. The Chan laboratory will employ MRM to evaluate specific changes in the mitochondrial proteome in which cycles of mitochondrial fission and fusion or mitophagy are disrupted. The Patterson lab will study a set of 14 target proteins in an established mouse model of maternal immune activation, a condition known to increase the risk of schizophrenia in the offspring of affected women by 3-7 fold. Insights in the molecular pathways that trigger this pathophysiology will suggest new targets for cures. The Varshavsky lab will quantify a set of previously defined substrates of the N-end rule pathway in budding yeast using MRM. This will provide molecular insight into the neurological deficits that result when the orthologous gene is mutated in humans. The availability of a state-of-the-art QTRAP 5500 mass spectrometer will enable these Caltech researchers to employ targeted MRM to quantitatively monitor changes in proteomes in response to a variety of chemical, genetic, and environmental perturbations. All of the projects relate to critical subcellular processes and many of the genes and proteins to be studied are targets of human therapeutics or are causative agents in human diseases including neurodegeneration, schizophrenia, and cancer. Thus, the proposed studies have the potential to have a great impact on human health.

描述（由申请人提供）：本申请是为了获得购买AB SCIEX QTRAP 5500 PRO三重四极杆（QQQ）质谱仪的资金，该质谱仪与Eksigent nanoLC相结合，用于进行基于多反应监测（MRM）的分析。MRM是一种非常敏感、定量和可重复的方法，用于监测复杂生物样品中靶向低丰度蛋白质的水平。从概念上讲，它类似于进行定量免疫印迹，但不需要标记或提高针对目标蛋白的抗体。从长远来看，该仪器将使许多以定量蛋白质科学为核心组成部分的研究项目成为可能。该应用程序包括八个具体项目。Deshaies实验室将量化酵母中cullin-RING泛素连接酶（CRL）网络中64种蛋白质的水平，这些蛋白质携带Nedd8/Rub1偶联酶和解偶联酶的突变。这一分析将提供对这一关键调控系统的分子逻辑的见解，该系统涉及癌症的发病机制。Rothenberg实验室将在小鼠身上研究一组明确的转录因子，这将为t细胞分化提供基本的见解。此外，戴维森实验室将研究一套明确的