CREATION OF A LEUKOCYTE PROTEOME MAP

白细胞蛋白质组图谱的创建

基本信息

批准号：
6400208
负责人：
IVAN C GERLING
金额：
$ 21.11万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-09 至 2004-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): It is proposed to develop a leukocyte proteome map as a new research resource for scientists studying immune mediated events in mice. The proteome consists of all of the proteins that are expressed at any one time, in a cell or tissue. A proteomic assay is, therefore, by definition the most encompassing assay for changes at the protein level. It is the protein equivalent of DNA microarray assays. By evaluating shifts in 2D gel electrophoresis (2DGE) proteome patterns of blood leukocytes, immunologists can quickly find out if any of thousands of proteins are expressed at different levels in two leukocyte samples. Unfortunately, if interesting differences are found the process of identifying what protein is in the spot that changed expression requires resources and expertise not available to most scientists. High sensitivity mass spectrometry (MS) is used to characterize such proteins, via the molecular weight (mw) of the intact protein, and via peptide mapping (mw and amino acid sequence of e.g. tryptic peptides from the protein) It is proposed to make 2DGE proteome assay of mouse leukocytes available to most scientists by establishing a 2DGE proteome reference map of mouse blood leukocytes. This will allow instant identification of a large number of protein spots on 2D gels, eliminating the need for MS analysis to characterize spots on the gels. The initial three year proposal will be used to create a "low detail" map and solve some important technical questions, that must be resolved before an application for creating a full scale detailed mouse leukocyte proteome map can be submitted. The proposal has the following Specific Aims: Specific Aim 1. To establish a 2DGE proteome reference map of mouse blood leukocytes that will allow an instant identification of a number of protein spots on 2D gels, eliminating the need for MS analysis to characterize those protein spots. Specific Aim 2. To evaluate the extent to which such maps are useful in mouse strains different from the one used to create the map. Polymorphism in immunologically important molecules may create pattern differences between mouse strains. The extent to which such differences will interfere with the use of a leukocyte proteome map created in another strain cannot presently be predicted. Specific Aim 3. To evaluate the extent to which a general mouse leukocyte map can be used to study proteomes of individual leukocyte subsets. Important immunological events may occur only in specific subsets of leukocytes, and presently, it cannot be predicted to what extent the presence of proteins from all other leukocyte subsets may interfere with the ability to detect such events.

描述（由申请人提供）：建议开发白细胞蛋白质组图谱，作为研究免疫的科学家的新研究资源小鼠中介导的事件。蛋白质组由所有蛋白质组成在细胞或组织中的任何时刻表达。蛋白质组学测定是，因此，根据定义，最全面的变化分析蛋白质水平。它相当于 DNA 微阵列检测的蛋白质。经过评估血液 2D 凝胶电泳 (2DGE) 蛋白质组模式的变化白细胞中，免疫学家可以快速找出数千种蛋白质中的任何一种在两个白细胞样本中以不同水平表达。不幸的是，如果在识别蛋白质的过程中发现了有趣的差异在改变表达方式需要资源和专业知识的地方，而不是大多数科学家都可以使用。使用高灵敏度质谱 (MS) 通过完整的分子量 (mw) 来表征此类蛋白质蛋白质，并通过肽图谱（分子量和氨基酸序列，例如胰蛋白酶蛋白质中的肽）建议对小鼠白细胞进行 2DGE 蛋白质组分析大多数科学家通过建立小鼠血液的 2DGE 蛋白质组参考图谱白细胞。这将允许立即识别大量 2D 凝胶上的蛋白质点，无需 MS 分析即可表征凝胶上有斑点。最初的三年提案将用于创建一个 “低细节”地图并解决一些重要的技术问题，这必须是在申请创建全尺寸详细鼠标之前已解决可以提交白细胞蛋白质组图谱。该提案有以下内容具体目标：具体目标 1. 建立 2DGE 蛋白质组参考图谱小鼠血液白细胞可以立即识别数字 2D 凝胶上的蛋白质斑点，无需 MS 分析表征这些蛋白质点。具体目标 2. 评估程度哪些图谱对于与过去使用的小鼠品系不同的小鼠品系有用创建地图。免疫学重要分子的多态性可能在小鼠品系之间产生模式差异。这种程度差异将干扰创建的白细胞蛋白质组图谱的使用目前无法预测另一种菌株。具体目标 3. 评估一般小鼠白细胞图谱可用于研究的程度单个白细胞亚群的蛋白质组。重要的免疫学事件可能仅发生在白细胞的特定亚群中，目前还无法确定预测来自所有其他白细胞的蛋白质的存在程度子集可能会干扰检测此类事件的能力。