5500 Q-Trap Mass Spectrometer

5500 Q-Trap 质谱仪

• 批准号: 7794200
• 负责人: STEPHEN BARNES
• 金额: $ 46.92万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2011-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794200
• 关键词: Age; Aging; Amino Acids; Area; Atherosclerosis; Biological; Cataract; Chronic Obstructive Airway Disease; Collagen; Complex; Crystallins; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Microarray Chip; Data; Diabetes Mellitus; Disease; Enzymes; Genes; Glycolysis; Ions; Length; Life; Light; Liquid substance; Lung; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Microarray Analysis; Mitochondria; Modeling; Monitor; Names; Organism; Oxidative Stress; Pathway interactions; Peptides; Plasma; Post-Translational Protein Processing; Preparation; Protein Chemistry; Proteins; Proteomics; Public Health; Reaction; Research; Research Personnel; Research Support; Role; Running; Scanning; Schedule; Speed; Stable Isotope Labeling; Time; Tissues; United States National Institutes of Health; Urine; base; biological systems; improved; instrument; interest; lens; mass spectrometer; methionyltryptophan; multicatalytic endopeptidase complex; neutrophil; protein complex; protein degradation; public health relevance; research study; tryptophyltyrosine

DESCRIPTION (provided by applicant): This is an application for an ABI-Sciex 5500 Qtrap to support research by 8 named NIH investigators who are carrying out research on biological pathways, protein complexes, protein post-translational modifications and small biologically active peptides in living systems. The 5500 Qtrap has significant sensitivity advantages over the existing 4000 Qtrap with a 5-fold increase in the multiple reaction ion monitoring (MRM) mode and >30 fold for MSMS analysis. The high sensitivity plus the very fast scanning speed (up to 20,000 m/z per sec) is a significant advance. It allows the monitoring of 100 MRM channels with the ability to capture a confirmatory MSMS spectrum during the passage of a peak in each MRM channel. With elution time scheduling, the number of channels monitored in a single LC run can be substantially increased. This instrument is well suited to directed proteomics in which the proteins of interest are well-defined, either by other protein mass spectrometry experiments, from DNA microarray analysis, gene sequencing, or from other biological results. In this approach, peptides representing proteins are carefully selected based on length (8-18 residues), amino acid composition (omitting peptides with easily oxidized residues - Cys, His, Met, Trp and Tyr) and sequence uniqueness to act as surrogates. These peptides can also be measured quantitatively by the preparation of stable isotope-labeled forms. This allows whole pathways of proteins/enzymes or protein complexes to be analyzed in a single LC run and hence to understand the role of protein turnover. These include the proteasome complex in activated neutrophils, the enzymes in glycolysis and energy generating mitochondrial pathways, and the lens crystallins in aging. The large number of MRM channels also allows the study of heavily modified proteins such as the cystic fibrosis transmembrane regulator (we have observed over 60 post- translational modifications) and lens crystallins. The high sensitivity of the 5500 Qtrap will also allow measurement of small peptides such as those produced from collagen in the lung in chronic obstructive pulmonary disease to be measured in more convenient/accessible fluids such plasma or urine. The key issue in all these experiments is that the investigators want to carry out analyses on proteins in biological systems at normal abundance levels. The increased sensitivity of the 5500 Qtrap significantly improves that capability. PUBLIC HEALTH RELEVANCE: The requested instrument is much more sensitive than its predecessors and will allow investigators to examine the complexity of protein chemistry in living tissues. It will be applied to the study of mechanisms of neutrophilassociated damage in the lung, mitochondrial-associated damage in models of oxidative stress (atherosclerosis, diabetes and cancer), lens cataract disease in aging, pathway robustness in the light of gene damage/loss, and in Cystic Fibrosis. It's very well suited to obtaining quantitative data to better understand the role of the protein(s) in specific areas of public health.

描述（由申请人提供）：这是一份ABI-Sciex 5500 Qtrap的申请，用于支持8名NIH研究人员的研究，这些研究人员正在进行生物学途径、蛋白质复合物、蛋白质翻译后修饰和生命系统中的小生物活性肽的研究。与现有的4000 Qtrap相比，5500 Qtrap具有显著的灵敏度优势，在多反应离子监测（MRM）模式下增加了5倍，在MSMS分析中增加了>30倍。高灵敏度加上非常快的扫描速度（高达每秒20，000 m/z）是一个重大的进步。它允许监测100个MRM通道，并能够在每个MRM通道中的峰通过期间捕获确证性MSMS光谱。利用洗脱时间调度，可以显著增加在单个LC运行中监测的通道数量。该仪器非常适合于定向蛋白质组学，其中感兴趣的蛋白质通过其他蛋白质质谱实验、DNA微阵列分析、基因测序或其他生物学结果得到明确定义。在该方法中，基于长度（8-18个残基）、氨基酸组成（省略具有容易氧化的残基- Cys、His、Met、Trp和Tyr的肽）和序列独特性仔细选择代表蛋白质的肽以充当替代物。这些肽也可以通过制备稳定的同位素标记形式来定量测量。这允许在单个LC运行中分析蛋白质/酶或蛋白质复合物的整个途径，从而了解蛋白质周转的作用。这些包括活化的中性粒细胞中的蛋白酶体复合物、糖酵解和能量产生线粒体途径中的酶以及老化中的透镜晶体蛋白。大量的MRM通道还允许研究严重修饰的蛋白质，例如囊性纤维化跨膜调节因子（我们已经观察到超过60种翻译后修饰）和透镜晶体蛋白。5500 Qtrap的高灵敏度还将允许在更方便/可获得的液体（如血浆或尿液）中测量小肽，如慢性阻塞性肺病患者肺中胶原蛋白产生的小肽。所有这些实验的关键问题是，研究人员希望在正常丰度水平下对生物系统中的蛋白质进行分析。5500 Qtrap灵敏度的提高显著提高了这一能力。 公共卫生关系：所要求的仪器比以前的仪器灵敏得多，将使研究人员能够检查活组织中蛋白质化学的复杂性。它将被应用于研究肺中嗜中性粒细胞相关损伤、氧化应激模型中的视网膜相关损伤（动脉粥样硬化、糖尿病和癌症）、衰老中的透镜白内障疾病、基因损伤/丢失中的通路稳健性以及囊性纤维化的机制。它非常适合获得定量数据，以更好地了解蛋白质在公共卫生特定领域的作用。