New proteome technologies: mapping adult D. melanogaster

描述(由申请人提供)：本提案描述了一项开发多维分析技术的计划，该技术用于分析从黑腹果蝇中提取的复杂生物混合物，作为生物体年龄的函数。初步数据表明，一种新的气相离子迁移率/飞行时间质谱学技术可以与其他分离技术相结合，以分离和表征从可溶蛋白质消化中获得的胰蛋白酶多肽的复杂混合物。气相分离减少了仅与质谱学相关的光谱拥堵和化学噪声，提供了关于电荷状态和氨基酸组成的额外信息，并允许使用一种并行方法来获得碎片数据(这是指定肽序列和识别蛋白质的重要因素)。本提案描述了这些技术的进一步发展及其在新兴蛋白质组学领域的应用。提出了一个激进的计划，旨在绘制基本模式果蝇生物体的蛋白质组图。这一系统的详细蛋白质组图的存在是一个重要的科学步骤，将使一系列遗传模型得到详细研究。选择这些模型是为了显示人类遗传学研究难以解决的疾病状态(例如，神经退行性疾病)的特征。

DESCRIPTION (provided by applicant): This proposal describes a program for developing multidimensional analytical techniques for the analysis of complex biological mixtures that are extracted from Drosophila melanogaster as a function of the age of the organism. Preliminary data show that a new gas-phase ion mobility/time-of-flight mass spectrometry technique can be coupled to other separation techniques in order to separate and characterize complex mixtures of tryptic peptides obtained from digestion of soluble proteins. The gas-phase separation reduces spectral congestion and chemical noise associated with mass spectrometry alone, provides additional information about charge state and amino acid composition, and allows a parallel approach for obtaining fragmentation data (an important factor for assigning peptide sequence and identifying proteins). This proposal describes the further advancement of these technologies and their applications to the emerging field of proteomics. An aggressive plan that aims to map the proteome for the fundamental model Drosophila organism is proposed. This existence of a detailed proteomic map for this system is an important scientific step that will allow a range of genetic models to be investigated in detail. These models are chosen to display characteristics of diseased states (e.g., neurodegenerative conditions) for which genetic studies in humans are intractable.