PROTEIN IDENTIFICATION CORE

蛋白质鉴定核心

• 批准号: 6846672
• 负责人: NEIL L KELLEHER
• 金额: $ 34.24万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6846672
• 关键词: bioinformatics; biological signal transduction; biomedical facility; cell cell interaction; drug abuse; mass spectrometry; matrix assisted laser desorption ionization; neurosciences; neurotoxicology; posttranslational modifications; protein localization; protein sequence; protein structure function; proteomics

The entire constituent of intracellular signaling molecules expressed in the organisms under investigation represents an extremely complex analytical challenge with a vast number of components varying dramatically in size and concentration and expressed in a highly heterogeneous manner within the CNS of these organisms. To address these challenges, and to provide our users with a battery of state-of-the-art protein identification techniques, this core proposes the use and development of techniques capable of molecular interrogation in three separate areas. We will (1) identify signaling molecules by a variety of "Bottom Up" approaches including mass fingerprinting, automated MS/MS sequencing, and accurate mass measurement, (2) characterize identified intercellular signaling molecules by intact protein analysis to identify post-translational modifications that are common among intercellular signaling molecules and correct for possible errors in the genomic databases due to signal peptide cleavage etc., and (3) localize intercellular signaling molecules by a variety of MS imaging techniques including current state-of-the-art and development of proposed "stretched sample" and "capillary array" methods that have the potential to enhance resolution and sensitivity of imaging methods to a level needed by our users. The needs for intercellular signaling molecule identification of our users are quite different from "shotgun" proteomics of unicellular organisms in sample size and complexity. With the combined expertise and facilities in the groups associated with the proposed center, a unique opportunity exists to identify and characterize intercellular signaling molecules at their sources.

在生物体中表达的细胞内信号分子的全部成分 调查代表了极其复杂的分析挑战，涉及大量数据 成分的大小和浓度变化很大，并以高度表达 这些生物体的中枢神经系统内的异质方式。为了应对这些挑战，并 为我们的用户提供一系列最先进的蛋白质鉴定技术，这个核心 提出使用和开发能够在三个方面进行分子询问的技术 单独的区域。我们将 (1) 通过各种“自下而上”的方法识别信号分子 包括质量指纹、自动 MS/MS 测序和精确质量测量，(2) 通过完整蛋白质分析来表征已识别的细胞间信号分子，以识别细胞间信号分子中常见的翻译后修饰，并纠正由于信号肽裂解等而导致的基因组数据库中可能存在的错误，以及 (3) 通过 各种 MS 成像技术，包括当前最先进的技术以及所提出的“拉伸样品”和“毛细管阵列”方法的开发，这些方法有可能将成像方法的分辨率和灵敏度提高到我们用户所需的水平。我们用户对细胞间信号分子鉴定的需求与单细胞生物的“鸟枪式”蛋白质组学在样本量和复杂性上有很大不同。凭借与拟议中心相关的小组的综合专业知识和设施，存在独特的机会来识别和表征细胞间信号分子的来源。