Development of New Proteomics Technology and its Application to Study Cellular Organization

蛋白质组学新技术的发展及其在细胞组织研究中的应用

• 批准号: 10607046
• 负责人: Martin Wühr
• 金额: $ 4.91万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607046
• 关键词: Affinity; Award; Cell Nucleus; Collaborations; Complement; Complex; Development; Developmental Biology; Drosophila genus; Embryo; Embryonic Development; Escherichia coli; Genetic Transcription; Goals; Importins; Improve Access; Ions; Measurement; Messenger RNA; Methods; Nuclear Proteins; Organism; Parents; Preparation; Printing; Protein Dynamics; Proteins; Proteomics; Publishing; Rana; Regulation; Reporter; System; Techniques; Technology; Translations; Work; Xenopus; Yeasts; cost; data integration; insight; instrument; mass spectrometer; multiple omics; protein degradation; technology development

Deciphering the Relationship Between mRNA and Protein Dynamics in a Developing Proto-Vertebrate The broad goal of the parent award (Development of New Proteomics Technology and its Application to Study Cellular Organization) is to advance quantitative proteomics technology and apply it to obtain a systems level understanding of cellular organization. Recently, we have demonstrated that TMTproC significantly increases sensitivity and multiplexing capacity of the complement reporter ion approach while making these approaches compatible with a broad set of mass spectrometers (Johnson, 2021). We have also worked to improve access to high- quality proteomics data by integrating TMTproC with ion-trap only instruments which cost 80% less than ones typically used for multiplexed quantitative proteomics. We have successfully leveraged our proteomics technology development for multiple collaborations (Crapse, 2021; Hart, 2020; Cao, 2020) which has advanced our understanding of cellular organization. One focus of the lab is to better understand developmental progression (Nguyen, 2021; Keber, 2021; Nofal, 2021). Recently, we have published a preprint showing that, in frog embryos, nuclear proteins access the nucleus in a temporal order set via importin affinities during development, thereby acting as an elegant timing mechanism to control the onset of their downstream functions (Nguyen, 2021). Extending beyond our work in developmental biology, we have developed methods that allow us to parameterize all aspects of protein abundance control on a global scale through multi-omics integration of transcription, translation, and degradation. Key to these advances was developing new methods that allow measurement of perturbation-free protein turnover. So far, we have applied these methods to E. coli and yeast (Gupta, et al., in preparation), and Alex Frese is expanding these approaches to the developing Ciona embryo with the goal of generalizing these techniques to complex higher organisms such as Xenopus and Drosophila.

解释发育过程中信使核糖核酸和蛋白质动力学的关系 原生脊椎动物 父奖项的广泛目标(新蛋白质组学技术及其发展 应用于细胞组织研究)是推进定量蛋白质组学技术的基础 并应用它来获得对蜂窝组织的系统层面的理解。最近，我们有 证明了TMTproC显著提高了信号的灵敏度和多路传输能力 补充报告方法，同时使这些方法与广泛的集合兼容 (约翰逊，2021年)。我们还努力改善获得高- 将TMTproC与仅需80%成本的离子陷阱仪器相集成，可获得高质量的蛋白质组学数据 少于通常用于多重定量蛋白质组学的那些。我们已经成功地 利用我们的蛋白质组学技术开发进行多项协作(Crapse，2021； Hart，2020；曹，2020)，这促进了我们对细胞组织的理解。一 该实验室的重点是更好地理解发育过程(Nguyen，2021；Keber，2021； 诺法尔，2021年)。最近，我们发布了一份预印本，显示在青蛙胚胎中，细胞核 蛋白质在发育过程中通过导入蛋白亲和力以时间顺序进入细胞核， 从而作为一种优雅的计时机制来控制其下游的开始 函数(阮氏，2021年)。除了我们在发育生物学方面的工作之外，我们还有 开发的方法使我们能够对蛋白质丰度控制的所有方面进行参数化 通过转录、翻译和降解的多组学集成实现全球规模。关键是 这些进展正在开发新的方法，使测量不受干扰 蛋白质周转率。到目前为止，我们已经将这些方法应用于大肠杆菌和酵母(Gupta等人，在 准备)，Alex Frese正在将这些方法扩展到正在发育的Ciona胚胎 目标是将这些技术推广到复杂的高等生物，如非洲爪哇 还有果蝇。