New proteome techniques: mapping adult D. Melanogaster

新的蛋白质组技术：绘制成年黑腹果蝇图谱

• 批准号: 9146961
• 负责人: DAVID E. CLEMMER
• 金额: $ 29.12万
• 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/9146961
• 关键词: Adult; Age; Alzheimer' s Disease; Animals; Antibodies; Area; Beds; Biological; 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; Health; 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; biomarker discovery; 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; 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（由申请人提供）：生物标志物发现工作的一个问题是当前的分析方法无法及时分析复杂生物样本中的众多分子种类。这个问题由于必须分析许多不同的样品以确定任何给定分子种类在群体中的正常变异性的事实而加剧。在这里，我们提出了一种新的分析仪器，利用快速，气相分离，以增加整体实验吞吐量超过一个数量级的发展。新仪器将采用多维离子迁移谱（IMS）技术，以获得快速分析。通过使用IMS分离的两个维度和改变诸如电场和测量温度的操作参数，预期将获得超过104的峰值容量（在质谱分析之前）。这是值得注意的，因为它比从二维液相色谱（LC）分离获得的更大，并且需要一小部分时间。新仪器将用于表征D。黑腹菌蛋白质组在生物体寿命的不同时期。显著增加的通量将允许将个体生物体与群体平均值进行比较，以根据分子表型更好地对生物体进行分组。此外，将采用一种新的荧光标记策略，以更好地比较个体生物体的生理年龄，而不是实足年龄。该测量平台将应用于转基因帕金森病和阿尔茨海默病果蝇，以确定与疾病进展相关的分子变化。