Proteome Map of the Photoreceptor Cell

描述（由申请人提供）：本提案的总体目标是探索眼部蛋白质组学方法的能力，以获得有关单个细胞类型及其单个亚细胞区室的蛋白质组成的新信息。具体目标是生成视网膜感光细胞层的蛋白质组图谱，其中大量的单个蛋白质将被分配到它们的相对亚细胞位置。这将通过在整个光感受器层的渐进切向切片中进行全面的质谱蛋白质测序来实现。数据将在数据库中汇总，其中所有鉴定的蛋白质将根据其亚细胞定位以及其在各种功能组中的潜在参与进行分组。获得这样一个蛋白质组图谱将使我们和许多其他研究人员能够完善现有的假设，并制定新的假设，广泛的感光功能和疾病。这项研究开发的技术也将作为类似研究的模板，解决视网膜其他层以及其他分层组织中的蛋白质分布。本申请中描述的项目实现了本计划公告（PAR-03-107）的目标，主要有两个原因。首先，它有可能导致一个重大的技术突破，建立方法进行大规模的眼组织蛋白质组学分析。随着人类进入后基因组时代，在眼科和视觉科学的许多领域迫切需要发展先进的蛋白质组学方法。据我们所知，所提出的研究的几个关键方面（例如连续组织切片的分析和“二维色谱”肽分离方案的使用）尚未用于应用于任何眼组织。其次，该项目旨在获得对视觉研究具有潜在高度影响的数据，因为单个感光细胞室的蛋白质组成的知识将使许多研究感光细胞生物学中广泛问题的科学家受益，并阐明眼部疾病的分子基础。

DESCRIPTION (provided by applicant): The overall objective of this proposal is to explore the powers of the ocular proteomics approach for obtaining novel information on protein composition of individual cell types and their individual subcellular compartments. The Specific Aim is to generate a proteome map of the photoreceptor cell layer of the retina where a large number of individual proteins will be assigned to their relative subcellular locations. This will be achieved by conducting comprehensive mass spectrometry protein sequencing in progressive tangential sections throughout the photoreceptor layer. The data will be summarized in a database where all identified proteins will be grouped according to their subcellular localization, as well as their potential involvement in various functional groups. Obtaining such a proteome map would enable us and many other investigators to refine existing hypotheses and to formulate novel hypotheses regarding a broad array of photoreceptor functions and diseases. The technologies developed in this study will also serve as a template for similar investigations addressing protein distributions in other layers of the retina as well as other layered tissues. The project described in this application fulfills the goals of this Program Announcement (PAR-03-107) for two major reasons. First, it has a potential to lead to a significant technological breakthrough by establishing the methodologies for conducting large scale proteomics analyses of ocular tissues. As we are entering the post-genomics era, there is a pressing need to developing advanced proteomics approaches in many areas of ophthalmology and visual sciences. To our knowledge, several key aspects of the proposed studies (such as the analysis of serial tissue sections and the use of "two-dimensional chromatography" peptide separation protocols) have not been used in application to any ocular tissue. Second, this project aims to acquire a body of data that has a potentially high-impact on vision research because knowledge of the protein composition of individual photoreceptor compartments will benefit many scientists studying a broad array of problems in photoreceptor cell biology and elucidating molecular bases of eye diseases.