Chemical tools to decode nuclear lamina-ome

解码核纤层组的化学工具

基本信息

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
项目类别：
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

项目摘要

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.

项目概要该应用的目标是对活细胞中的核层相互作用蛋白进行成像和识别。这核层位于内核膜下方，由四个主要同源结构组成蛋白质，核纤层蛋白 A (LA)、核纤层蛋白 B1 (LB1)、核纤层蛋白 B2 (LB2) 和核纤层蛋白 C (LC)。层粘连蛋白是 V 型中间体丝蛋白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被显着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。核纤层蛋白在这些中的功能该过程是由复杂但不完全了解的核内蛋白质-蛋白质相互作用介导的叶片。更多的事实进一步强调了 LA 在这些生物过程中的重要性已知超过 450 个 LA 突变可引起多种疾病，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和在生理相关条件下对活细胞中的层结合蛋白进行成像和识别的技术状况。我们最近开发了一种名为 LBL1 的小分子，可以特异性结合整个核纤层蛋白。细胞蛋白质组。我们对 LBL1 的鉴定为我们提供了前所未有的机会来成像和鉴定活细胞中的内源核纤层蛋白结合蛋白。这种理解将会产生新颖的对核纤层蛋白功能的深入了解，并可能为开发治疗方法提供新途径核纤层蛋白病。为实现这一目标，我们提出以下两个具体目标： 1）开发用于标记活细胞核纤层的创新化学探针； 2) 识别和成像 LA 及其活细胞中相互作用的伙伴。