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ABI-SCIEX 5500 QTRAP的申请，以支持8名NIH研究者的研究，他们正在对生物途径，蛋白质复合物，蛋白质复合物，翻译后蛋白质后修饰和生物学中的小生物学活性肽进行研究。 5500 QTRAP比现有4000 QTRAP具有显着的灵敏度优势，而多反应离子监测模式（MRM）模式增加了5倍，MSMS分析> 30倍。高灵敏度加上非常快速的扫描速度（每秒最高20,000 m/z）是一个重大进步。它允许监视100个MRM通道，并能够在每个MRM通道中的峰通过期间捕获确认性MSMS频谱。通过洗脱时间调度，可以大大增加单个LC运行中监测的通道数。该仪器非常适合定向蛋白质组学，其中其他蛋白质质谱实验可以从DNA微阵列分析，基因测序或其他生物学结果中获得明确定义的蛋白质。在这种方法中，根据长度（8-18个残基），氨基酸组成（省略具有易于氧化的残基的肽 - Cys，His，Met，TRP和Tyr）和序列独特性作为代替代物的替代性。这些肽也可以通过制备稳定的同位素标记的形式进行定量测量。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，从而了解蛋白质更新的作用。其中包括活化的嗜中性粒细胞中的蛋白酶体复合物，糖酵解和产生线粒体途径的能量的酶，以及衰老中的透镜晶体。大量的MRM通道还允许研究重量修饰的蛋白质，例如囊性纤维化跨膜调节剂（我们已经观察到60多次翻译后修饰）和晶状体结晶蛋白。 5500 QTRAP的高灵敏度还将允许测量小肽，例如在慢性阻塞性肺疾病中肺中胶原蛋白产生的肽，以更方便/可及的液体（如血浆或尿液）进行测量。所有这些实验的关键问题是，研究人员希望在正常的丰度水平上对生物系统中的蛋白质进行分析。 5500 QTRAP的灵敏度的提高显着提高了该能力。 公共卫生相关性：所请求的仪器比其前辈更敏感，并将允许研究人员检查活组织中蛋白质化学的复杂性。它将应用于肺中性粒细胞相关损伤的机制，在氧化应激模型（动脉粥样硬化，糖尿病和癌症）中与线粒体相关的损伤，衰老中的透镜透镜疾病，基因损伤/囊肿纤维化中的途径稳健性。它非常适合获取定量数据，以更好地了解蛋白质在特定公共卫生领域的作用。