Proteomics Core

B1.客观化 蛋白质组学核心旨在为儿童/哈佛IDDRC的所有研究人员提供最先进的基于质谱学的蛋白质组学，以推进神经发育障碍的细胞和分子生物学研究。这一核心为蛋白质的质谱学分析提供指导、咨询和服务，包括蛋白质鉴定、翻译后修饰的表征以及具有广泛复杂性的样品中蛋白质的定量。 概述。基于质谱学的蛋白质组学是目前可用于表征蛋白质的最灵敏、定量和全面的技术。核心B的一个主要目标是使用蛋白质组学来帮助IDDRC研究人员识别与神经元功能相关的蛋白质复合体，测量神经系统在发育过程中和在导致发育障碍的条件下发生的蛋白质组成的数量变化，并确定神经元蛋白质的功能性翻译后修饰。 蛋白质组学是一个快速发展的领域，核心致力于在新的方法和应用出现时引入和实施它们。斯蒂恩博士是儿童医院神经生物学项目的助理教授，也是蛋白质生物化学和质谱学方面的专家。他应聘成立了IDDRC蛋白质组学核心，现在担任位于儿童柯比神经生物学中心的IDDRC蛋白质组学核心的主任。斯蒂恩实验室积极参与蛋白质组学研究，并设有一个计算/生物信息学小组，开发与蛋白质组学相关的统计和数据分析工具，并通过核心向IDDRC的所有成员提供使用这种统计和计算专业知识的途径。该中心为IDDRC成员提供蛋白质定量和比较蛋白质组学方面的建议和技术。 该核心旨在提供定性和定量的基于质谱学的蛋白质组学技术。自2005年蛋白质组学核心成立以来，这些强大的蛋白质组技术已成功地在IDDRC的许多实验室用于细胞和组织的蛋白质组分析，以研究智力和发育疾病。 蛋白质组学技术在大脑发育研究中的应用将完成：1)确定细胞和组织中蛋白质的完整星座的特征；2)作为疾病或细胞状态变化的功能，评估蛋白质含量或修饰的变化；以及3)蛋白质功能的特征。这些研究既包括大规模的蛋白质组实验，也包括定向的蛋白质组学实验。大规模的研究往往是定量的，使用同位素标记方法来寻找差异表达或修饰的蛋白质；而定向研究则检查单个蛋白质的结构，包括它与蛋白质功能有关的共翻译和翻译后修饰。蛋白质组学核心提供涵盖这两个领域的服务。 基因组测序提供了使用以前不可能的方法来解决有关大脑发育、疾病和功能障碍的问题的机会。为了了解智力发育和病理生理学的分子机制，IDDRC的研究人员现在可以直接询问细胞或组织的“蛋白质组”。该核心将通过提供蛋白质鉴定服务、翻译后修饰分析和蛋白质定量，为IDDRC研究人员提供一个重要的互动中心。

B1. OBJECTIVE The Proteomics Core aims to provide all investigators at the Children's/Harvard IDDRC with access to the state-of-the-art mass spectrometry-based proteomics required to advance studies in the cell and molecular biology of neurodevelopmental disabilities. This core provides instruction, consultation and service for the analysis of proteins by mass spectrometry, including protein identification, characterization of post-translational modifications, and quantification of proteins in samples with a wide range of complexities. Overview. Mass spectrometry-based proteomics is currently the most sensitive, quantitative and comprehensive technology available for the characterization of proteins. A major goal of Core B is to use proteomics to assist IDDRC investigators in the identification of protein complexes relevant to neuronal function, to measure the quantitative changes in protein composition that occur in the nervous system during development and under conditions that lead to developmental disabilities, and to identify functional posttranslational modifications of neuronal proteins. Proteomics is a rapidly progressing field and the Core is committed to introducing and implementing new methodology and applications as they become available. Dr. Steen, an Assistant Professor in the Neurobiology Program at Children's Hospital, and an expert in protein biochemistry and mass spectrometry, was recruited to setup and now act as director of the IDDRC Proteomics Core located within the Kirby Neurobiology Center at Children's. The Steen Laboratory is actively involved in proteomics research and has a computational/bioinformatics subgroup, which develops proteomics related statistical and data analysis tools and provides access to this statistical and computational expertise via the Core to all members of the IDDRC. The Core provides advice and the technology for protein quantification and comparative proteomics to IDDRC members. This Core has been designed to provide qualitative and quantitative mass spectrometry-based proteomics technologies. Since the inception of the Proteomics Core in 2005, these powerful proteomic technologies have been successfully used in many IDDRC laboratories for the proteomic analysis of cells and tissues to study intellectual and developmental diseases. The application of proteomic techniques to the study of the developing brain is poised to accomplish i) characterization of the full constellation of proteins in defined cells and tissues, ii) an assessment of changes in protein content or modifications as a function of disease or change in cell state, and iii) characterization of protein function. These investigations involve both large scale and directed proteomic experiments. Large scale studies are often quantitative, involving isotope tagging methods to find differentially expressed or modified proteins; whereas directed studies examine the structure of a single protein including its co-translational and post-translational modifications with respect to the protein's function. The proteomics Core provides services which encompass both these areas. Genome sequencing has provided the opportunity to address questions regarding brain development, disease and dysfunction using approaches previously impossible. To understand the molecular mechanisms of intellectual development and pathophysiology, IDDRC investigators can now directly interrogate the "proteome" of cells or tissues. This Core will provide a critical interactive center for IDDRC investigators by providing protein identification services, post-translational modification analyses and protein quantification.