Instrumentation Development: MS Array for Quantitative Proteomics

描述（由申请人提供）：基因组测序技术的快速发展使人类疾病在分子水平上的研究受益匪浅。然而，有效利用当前和未来基因组信息的主要障碍与基因功能分配固有的主要挑战有关。由于蛋白质是基因表达的主要功能结果，蛋白质组学领域的出现有助于解决这一需求。蛋白质组学含量的测量，包括蛋白质表达水平、翻译后修饰和蛋白质相互作用，可以产生与基因功能相关的关键见解。蛋白质组学研究的主要挑战与样品的复杂性和测量必须执行的动态范围有关。一般来说，这些需求比基因组学中遇到的要大得多，因为蛋白质丰度的变化比基因大得多，蛋白质在物理性质上比基因有更广泛的多样性，蛋白质没有信号放大的手段，也没有类似的沃森-克里克碱基配对。因此，蛋白质组学领域主要采用基于肽质谱测量的技术，因为这些测量已经显示出大规模蛋白质鉴定和定量的能力。然而，由于每个蛋白质平均可以产生50-100个肽，肽混合物的测量要比蛋白质的测量复杂得多。此外，目前依赖于数据的测量策略导致可实现动态范围的显著压缩，因为这种MS/MS测量通常只适用于观察到较高丰度的肽。理想情况下，蛋白质组消化中的每个肽都将进行质谱/质谱分析，以从蛋白质组学实验中获得最大的信息。该项目将通过开发一种质谱仪阵列来提高大规模蛋白质组学的能力，这种质谱仪阵列能够比目前最先进的技术更快地进行MS/MS采集。在这个项目下开发的质谱阵列技术将涉及离子回旋共振质谱，它将产生更高的质量分辨能力、更高的质量测量精度以及更高的吞吐量采集。因此，在一个给定的实验中可以识别一个数量级或更多的肽，这将极大地增加每个蛋白质组学分析的信息含量以及可以研究的蛋白质的动态范围。

DESCRIPTION (provided by applicant): The study of human diseases at the molecular level has benefitted greatly by rapid advances in technology for genome sequencing. However, major impediments for effective utilization of current and future genomic information relates to the major challenges that are inherent in functional assignment of genes. The field of proteomics has arisen to help address this need, since proteins are the predominant functional outcome of gene expression. Measurement of proteomic content, including protein expression levels, posttranslational modifications and protein interactions can yield critical insight relevant to gene function. The major challenges in proteomics research relate to the complexity of samples and the dynamic range over which measurements must be performed. In general terms, these demands are far greater than encountered in genomics since, protein abundances can vary much more than gene do, proteins have much wider diversity in physical properties than genes do, and proteins have no means for signal amplification, nor analogous Watson-Crick base pairing as genes do. Thus the field of proteomics employs technology largely based on mass spectrometry measurements of peptides since these measurements have shown capabilities for large-scale protein identification and quantitation. However, since each protein can produce on average, 50-100 peptides, measurements of peptide mixtures are far more complex than measurements on proteins. In addition, current data-dependent measurement strategies lead to significant compression of achievable dynamic range, since such MS/MS measurements are only normally feasible on peptides observed with higher abundance. Ideally, every peptide in a proteome-wide digest would be subjected to MS/MS to gain maximal information from proteomics experiments. This project will advance capabilities for large-scale proteomics through the development of a mass spectrometer array capable of MS/MS acquisition an order of magnitude faster than current state-of-the art technology. The MS array technology to be developed under this project will involve ion cyclotron resonance mass spectrometry that will yield higher mass resolving power, higher mass measurement accuracy as well as higher throughput acquisition. As a result, an order of magnitude or more peptides can be identified during a given experiment which will dramatically increase the information content of each proteomics analysis as well as the dynamic range of proteins that can be studied.