New proteome techniques: mapping adult D. Melanogaster

新蛋白质组技术：绘制成年黑腹果蝇图谱

• 批准号: 9009178
• 负责人: DAVID E. CLEMMER
• 金额: $ 25.41万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-22 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9009178
• 关键词: Adult; Age; Alzheimer' s Disease; Animals; Antibodies; Area; Beds; Biological; Biological Markers; Biological Models; Brain; Central Nervous System Diseases; Cessation of life; Comb animal structure; Complex; Complex Mixtures; DNA Sequence Alteration; Detection; Development; Dimensions; Disease Progression; Disease model; Drosophila genus; Drosophila melanogaster; Fluorescence; Gases; Gene Expression; Genes; Genome; Genotype; Human; Huntington Disease; Individual; Ions; Isomerism; Label; Liquid Chromatography; Longevity; Maps; Mass Spectrum Analysis; Measurement; Measures; Modeling; Molecular; Molecular Chaperones; Monitor; Nature; Nerve Degeneration; Onset of illness; Organism; Parkinson Disease; Pathway interactions; Peptides; Phase; Physiological; Polysaccharides; Population; Population Analysis; Population Control; Positioning Attribute; Program Development; Protein Conformation; Protein Isoforms; Proteins; Proteome; Proteomics; Reproducibility; Research; Resolution; Sampling; Source; Spectrometry; Speed; Structure; Study models; System; Techniques; Technology; Temperature; Testing; Time; Transgenic Organisms; Tube; Validation; Work; age related; analytical method; comparative; electric field; genetic strain; genetic variant; high throughput analysis; improved; instrument; instrumentation; ion mobility; molecular phenotype; mutant; new technology; next generation; novel; parkin gene/protein; protein complex; protein expression; public health relevance; research study; small molecule; synuclein; tool; two-dimensional

 DESCRIPTION (provided by applicant): A problem of biomarker discovery efforts is that current analytical methods do not have the capacity to analyze in a timely manner the numerous molecular species within complex biological samples. This problem is exacerbated by the fact that many different samples must be analyzed in order to establish the normal variability across a population for any given molecular species. Here we propose the development of a new analytical instrument that utilizes rapid, gas-phase separations to increase the overall experimental throughput by more than an order of magnitude. The new instrumentation will employ multidimensional ion mobility spectrometry (IMS) techniques to obtain the rapid analysis. By using two dimensions of IMS separations and changing operational parameters such as the electric field and measurement temperature it is anticipated that peak capacities (prior to mass spectrometric analysis) in excess of 104 will be obtained. This is remarkable because it is larger than that obtained from two-dimensional liquid chromatography (LC) separations and requires a fraction of the time. The new instrumentation will be used to characterize the D. melanogaster proteome at different times in the organism lifespan. The significantly increased throughput will allow comparisons of individual organisms with the population mean to better group organisms according to molecular phenotype. Additionally, a novel fluorescent labeling strategy will be employed to better compare the physiological age of individual organisms as opposed to chronological age. The measurement platform will be applied to transgenic Parkinson's and Alzheimer's disease Drosophila in order to determine molecular changes associated with disease progression.

 描述（由适用提供）：生物标志物发现工作的问题是，当前的分析方法没有能力及时分析复杂的生物样品中的许多分子物种。由于必须分析许多不同的样本，以便为任何给定的分子物种建立正常的变异性，这使这个问题加剧了这一事实。在这里，我们提出了一种新的分析仪器的开发，该仪器利用快速的气相分离将整体实验吞吐量提高到超过一个数量级。新仪器将采用多维离子迁移率（IMS）技术来获得快速分析。通过使用IMS分离的两个维度和更改的操作参数，例如电场和测量温度，可以预计，峰值容量（质谱分析之前）超过104，这是显着的，因为它比从二维液体色谱（LC）分离中获得的峰值容量要大，并且需要时间。新仪器将用于表征生物体寿命的不同时间D. melanogaster蛋白质组。吞吐量的显着增加将使单个生物体与种群的比较是根据分子表型与更好的组生​​物的比较。此外，将聘请一种新型的荧光标签策略，以更好地比较单个组织的身体时代，而不是按年代年龄进行比较。测量平台将应用于转基因帕金森氏病和阿尔茨海默氏病果蝇，以确定与疾病进展相关的分子变化。