Core A: Advanced Proteomics Core

项目摘要/摘要 核心A-高级蛋白质组学核心 核心A的目标是为全球和靶向提供基于质谱学的蛋白质组平台 转录调控因子在心脏发育和先天性心脏病中发挥核心作用的特征 心脏缺陷，以支持拟议的PPG项目1-3的目标和努力。我们是 不断开发新的和创新的策略，以实现更灵敏和定量的蛋白质分析- 蛋白质相互作用(PPI)和翻译后修饰(PTM)。这些努力现在让我们能够 超越静态的PPI或PTM，转向不同背景下的更动态的表征 发育阶段、基因敲除或基因突变。我们的亲和纯化质谱仪(AP-MS) 与新开发的基于邻近的生物素化(APEX-MS)一起的能力将有助于项目研究 1-3通过GATA4、TBX5、GATA4、TBX5、 MEF2C、Myocardin和BAF复合体成员。此外，我们的定量分析管道，以确定PTM 将为项目3中的研究人员提供直接支持，重点是MEF2C和Myocardin， 两个信号响应因素。所有这些实验都将在心脏相关系统中进行 诱导多能干细胞(IPSC)或胚胎干细胞(ESC)来源的心脏发育 前体细胞(CP)以及来自新生小鼠的心肌细胞(CMS)或心脏组织。超越 在这些项目的支持下，核心A将与核心B密切合作，为APEX-MS数据制定分析战略。 提供的专业知识、高灵敏度和定量的MS分析以及持续的创新 核心A将服务和适应项目和PPG项目核心的需要。

PROJECT SUMMARY/ABSTRACT CORE A – ADVANCED PROTEOMICS CORE The goal of Core A is to provide mass spectrometry–based proteomic platforms for global and targeted characterization of transcriptional regulators that play a central role in cardiac development and congenital heart defects in support of the aims and efforts for Projects 1–3 of the proposed PPG Project. We are continuously developing new and innovative strategies for more sensitive and quantitative analysis of protein- protein interactions (PPIs) and post-translational modifications (PTMs). These efforts now allow us to look beyond static PPIs or PTMs and towards more dynamic characterization in the context of different developmental stages, gene knockouts or gene mutations. Our affinity-purification mass spectrometry (AP-MS) capabilities along with newly developed proximity-based biotinylation (APEX-MS), will aid research in Projects 1–3 through dynamic interaction network mapping for transcriptional regulators, such as GATA4, TBX5, MEF2C, Myocardin and BAF complex members. Also, our quantitative analyses pipeline to determine PTMs on specific proteins will provide direct support for researchers in Project 3, focused on MEF2C and Myocardin, two signal responsive factors. All of these experiments will be performed in relevant systems for cardiac development, either induced pluripotent stem cell (iPSC)- or embryonic stem cell (ESC)-derived cardiac precursors (CPs), as well as cardiomyocytes (CMs) or cardiac tissue derived from neonatal mice. Beyond support of the projects, Core A will work closely with Core B to develop analysis strategies for APEX-MS data. The expertise, the highly sensitive and quantitative MS analysis, as well as ongoing innovations provided by Core A will serve and adapt to the needs of Projects and Cores of the PPG project.