Proteomics Core

B1.目的 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。该核心为通过质谱分析蛋白质提供指导，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。 概况.基于质谱的蛋白质组学是目前可用于表征蛋白质的最灵敏、最定量和最全面的技术。核心B的一个主要目标是利用蛋白质组学来协助IDDRC研究人员鉴定与神经元功能相关的蛋白质复合物，测量在发育过程中和导致发育障碍的条件下神经系统中发生的蛋白质组成的定量变化，并鉴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，核心致力于引入和实施新的方法和应用，因为它们变得可用。Steen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分析的专家，他被招募来建立IDDRC蛋白质组学核心，现在担任儿童医院Kirby神经生物学中心的主任。Steen实验室积极参与蛋白质组学研究，并拥有一个计算/生物信息学小组，该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的所有成员提供这种统计和计算专业知识。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。 该核心旨在提供基于质谱的定性和定量蛋白质组学技术。自2005年蛋白质组学核心成立以来，这些强大的蛋白质组学技术已成功用于许多IDDRC实验室，用于细胞和组织的蛋白质组学分析，以研究智力和发育疾病。 将蛋白质组学技术应用于发育中的脑的研究准备完成i）在限定的细胞和组织中蛋白质的完整星座的表征，ii）作为疾病或细胞状态变化的函数的蛋白质含量或修饰的变化的评估，以及iii）蛋白质功能的表征。这些研究涉及大规模和定向蛋白质组学实验。大规模的研究通常是定量的，涉及同位素标记方法以找到差异表达或修饰的蛋白质;而定向研究检查单个蛋白质的结构，包括其与蛋白质功能相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。 基因组测序提供了机会，解决有关大脑发育，疾病和功能障碍的问题，使用以前不可能的方法。为了了解智力发育和病理生理学的分子机制，IDDRC研究人员现在可以直接询问细胞或组织的“蛋白质组”。该中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研究人员提供一个关键的互动中心。

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.