MASS SPECTROSCOPY/PROTEOMICS

质谱/蛋白质组学

• 批准号: 7414793
• 负责人: PETER D. EMANUEL
• 金额: $ 29.63万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414793
• 关键词: Area; Bioinformatics; Biometry; Cells; Collaborations; Coomassie blue; Cyclotrons; Detection; Dyes; Endopeptidases; Faculty; Fingerprint; Fourier Transform; Funding; Future; Gel; Grant; Hybrids; Image; Image Analysis; Ions; Label; Lasers; Liquid substance; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Methods; National Center for Research Resources; Operative Surgical Procedures; Peptide Hydrolases; Peptides; Physiological; Post-Translational Protein Processing; Procedures; Process; Protein Analysis; Protein Chemistry; Protein Sequence Analysis; Proteins; Proteome; Proteomics; Pump; Recovery; Research Personnel; Robot; Robotics; Sampling; Services; Solutions; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Spottings; Staining method; Stains; System; Therapeutic; Time; Tissue Sample; Tyrosine; University of Alabama at Birmingham Cancer Center; Xenobiotics; day; gel electrophoresis; instrument; mass spectrometer; microbial alkaline proteinase inhibitor; nano; nitration; programs; protein expression; protein protein interaction; protein structure; small molecule; tandem mass spectrometry

DESCRIPTION (provided by applicant): The Mass Spectrometry/Proteomics Shared Facility is organized to provide a variety of analytical and technical services using mass spectrometry to UAB Comprehensive Cancer Center investigators. The Shared Facility currently has five mass spectrometers - two MALD1-TOF instruments, two triple quadrupole instruments, and one hybrid quadrupole orthogonal TOF instrument. Three new instruments (a new triple quadrupole instrument and two fourier transform-ion cyclotron resonance instruments - supported by NCRR grants) will be installed during the summer of 2004. Besides qualitative and quantitative analysis of small molecules (therapeutic and xenobiotics) and peptides, the mass spectrometry group provides a variety of analyses in the area of protein chemistry and protein expression. These include H-D exchange to probe protein structure in solution as well as protein-protein interactions. Procedures have also recently been added to allow investigators to pursue proteomics applications. The proteomics section of the Shared Facility will provide investigators with consultant advice as how to prepare samples for analysis, and in most cases to carry out the analyses. This includes recovery of the proteome from the cell or tissue samples, resolving proteins using 1D-SDS-PAGE or 2D-IEF/SDS-PAGE, staining and imaging the gels, and carrying out sophisticated image analysis to detect changes that have occurred in the observable proteome. These are verified by statistical analysis by the Biostatistics/Bioinformatics Shared Facility. Protein spots are removed robotically and following robotic processing to generate a protease-induced peptide profile analysis by MALDI-TOF mass spectrometry. The peptide mass fingerprint is subjected to statistical analysis with the MASCOT search engine to putatively identify the protein. Verification of protein identity is performed by nano-LC tandem mass spectrometry of the peptides. This method also allows for the detection of post-translational modifications.

描述(申请人提供)：质谱学/蛋白质组学共享设施是 组织为UAB综合癌症中心的研究人员提供各种使用质谱学的分析和技术服务。共享设施目前有五个质量 光谱仪-两台MALD1-TOF仪器，两台三重四极仪器和一台混合式仪器 四极正交飞行时间测定仪。将在2004年夏季安装三台新仪器(一台新的三重四极仪器和两台傅里叶变换离子回旋共振仪器--由NCRR赠款资助)。除了对小分子(治疗性和外源性)和多肽进行定性和定量分析外，质谱组还在蛋白质化学和蛋白质表达领域提供各种分析。其中包括H-D交换以探测溶液中的蛋白质结构以及蛋白质与蛋白质的相互作用。最近还增加了一些程序，允许调查人员进行蛋白质组学应用。共享设施的蛋白质组学部分将就如何准备样本进行分析以及在大多数情况下如何进行分析向调查人员提供咨询意见。这包括从细胞或组织样本中回收蛋白质组，使用1D-SDS-PAGE或2D-IEF/SDS-PAGE分解蛋白质，对凝胶进行染色和成像，以及执行复杂的图像分析以检测可观察到的变化 蛋白质组。这些都通过生物统计/生物信息学共享设施的统计分析进行了验证。机器人去除蛋白质斑点，然后进行机器人处理，以产生由MALDI-TOF质谱仪进行的蛋白酶诱导的多肽图谱分析。用吉祥物搜索引擎对多肽质量指纹图谱进行统计分析，以推测鉴定蛋白质。通过多肽的纳米LC串联质谱仪进行蛋白质同一性的验证。该方法还允许检测翻译后修饰。