Chemical tools to decode nuclear lamina-ome

解码核纤层组的化学工具

• 批准号: 10375520
• 负责人: Bingbing Li
• 金额: $ 23.1万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375520
• 关键词: Address; Award; Binding; Binding Proteins; Biological Process; Biotin; Buffers; CCI-779; Cell Nucleus; Cells; Cellular biology; Chemicals; Chromatin; Co-Immunoprecipitations; Complex; Copper; DNA Repair; Disease; Genetic Transcription; Goals; Homologous Protein; Human; Image; Intermediate Filament Proteins; Label; Lamin B1; Lamin Type A; Lamins; Ligand Binding; Ligase; Ligation; Mass Spectrum Analysis; Mechanics; Mediating; Methods; Morphologic artifacts; Mutation; National Institute of Biomedical Imaging and Bioengineering; Nuclear Envelope; Nuclear Inner Membrane; Nuclear Lamin; Nuclear Lamina; Nucleoplasm; Pathologic; Peroxidases; Phenols; Physiological; Process; Proteins; Proteome; Reaction; Regulation; Research; Research Personnel; Role; Set protein; Specificity; Streptavidin; Technology; ascorbate; base; crosslink; innovation; insight; lamin B2; lamin C; mechanotransduction; novel; novel therapeutics; overexpression; prevent; protein protein interaction; response; scaffold; small molecule; stem cells; therapy development; tool

Project Summary The goal of this application is to image and identify nuclear lamina-interacting proteins in living cells. The nuclear lamina, located underneath of inner nuclear envelope, is composed of four major homologous proteins, lamin A (LA), lamin B1 (LB1), lamin B2 (LB2) and lamin C (LC). Lamins are type V intermediate filament proteins and provide mechanical support for the mammalian nucleus. However, the roles of lamins have been significant extended into other critical aspects of cell biology including mechanosensing DNA repair, chromatin regulation, gene transcription and stem cell regulation. The function of lamins in these processes are mediated by complex yet incompletely understood protein-protein interactions at the nuclear lamina. The importance of LA in these biological processes is further emphasized by the fact that more than 450 LA mutations are known to cause a wide spectrum of diseases collectively called laminopathies. One of the major obstacles in addressing this challenge is the lack of the appropriate tools and technologies to image and identify the lamina-binding proteins in living cells under physiologically relevant conditions. We recently developed a small molecule called LBL1 that specifically binds lamins in the whole cellular proteome. Our identification of LBL1 provides an unprecedented opportunity for us to image and identify the endogenous lamin-binding proteins in living cells. This understanding will generate novel insights into the lamin functions and potentially provide novel avenues to develop therapies for laminopathies. To achieve this goal, we propose the following two specific aims: 1) To develop an innovative chemical probe to label nuclear lamina in living cells; 2) To identify and image LA and its interacting partners in living cells.

项目摘要 该应用程序的目标是成像和识别活细胞中的核纤层相互作用蛋白。的 核纤层位于内核膜的下方，由四个主要的同源核纤层组成， 核纤层蛋白A（LA）、核纤层蛋白B1（LB 1）、核纤层蛋白B2（LB 2）和核纤层蛋白C（LC）。核纤层是V型中间体 丝蛋白质，并为哺乳动物细胞核提供机械支持。然而，核纤层蛋白的作用 已经显著扩展到细胞生物学的其他关键方面，包括机械传感DNA 修复、染色质调节、基因转录和干细胞调节。核纤层蛋白在这些 这些过程是由复杂但尚未完全了解的蛋白质-蛋白质相互作用介导的 层。LA在这些生物过程中的重要性进一步强调了以下事实： 已知超过450种LA突变引起广泛的疾病，统称为核纤层蛋白病。 应对这一挑战的主要障碍之一是缺乏适当的工具， 在生理相关条件下对活细胞中的纤层结合蛋白进行成像和鉴定的技术 条件我们最近开发了一种名为LBL 1的小分子，它特异性地结合整个细胞中的核纤层蛋白。 细胞蛋白质组我们对LBL 1的鉴定为我们提供了一个前所未有的机会， 鉴定活细胞中的内源性层粘连蛋白结合蛋白。这种理解将产生新的 深入了解核纤层蛋白的功能，并可能提供新的途径来开发治疗， 核纤层蛋白病为达致这个目标，我们提出以下两个具体目标： 创新的化学探针标记活细胞中的核纤层; 2）识别和成像LA及其 在活细胞中相互作用的伙伴。