ROLE OF MECHANICAL FORCE IN NOTCH

机械力在 NOTCH 中的作用

• 批准号: 8362716
• 负责人: ELIAS BOTVINICK
• 金额: $ 1.56万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362716
• 关键词: Address; Adhesions; Adult; Affinity; Avidity; Binding; Biotechnology; Cell Culture Techniques; Cells; Cessation of life; Dissociation; Embryonic Development; Endocytosis; Funding; Generations; Grant; Lasers; Ligand Binding; Ligands; Maintenance; Mammalian Cell; Measures; Mechanics; Modeling; Morphogenesis; National Center for Research Resources; Pathway interactions; Pattern; Principal Investigator; Proteolysis; Publishing; Recycling; Research; Research Infrastructure; Resources; Role; Signal Transduction; Site; Source; Stem cells; System; Tissues; United States National Institutes of Health; Work; base; cell type; cost; laser tweezer; notch protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Notch pathway is an evolutionary conserved signaling system used extensively throughout embryonic development that continues to function in the maintenance of tissues and stem cells in adults. Notch signaling directs the specification of a variety of cell types and contributes to tissue patterning, and morphogenesis through effects on cellular differentiation, proliferation, survival and death. Our working model for ligand-induced Notch signaling is based on Weinmaster's published and preliminary findings obtained with mammalian cell culture, and incorporates the critical role for forces generated during ligand endocytosis with Notch structural findings. Specifically, we hypothesize that endocytosis of ligand-bound Notch generates a robust mechanical force that physically pulls the Notch receptor apart to expose the ADAM cleavage site and facilitate activating proteolysis. Our model also proposes that furin cleavage to produce the Notch heterodimer predisposes Notch to physical dissociation through mechanical force generated during ligand endocytosis. However, ligand endocytosis has also been proposed to facilitate recycling to generate a high affinity Notch ligand. To address this controversy, we are using optical tweezers to determine roles for ligand endocytosis in regulating binding strength and force generation. SMM laser tweezers used to measure endocytic forces and avidity between Notch coated beads and ligand expressing cells. Correlations between endocytic forces, adhesion strength and Notch transedocytoiss are sought.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 Notch 通路是一种进化保守的信号系统，在整个胚胎发育过程中广泛使用，在成人组织和干细胞的维持中继续发挥作用。 Notch 信号传导指导多种细胞类型的规范，并通过影响细胞分化、增殖、存活和死亡来促进组织模式和形态发生。我们的配体诱导Notch信号传导工作模型基于Weinmaster发表的哺乳动物细胞培养初步研究结果，并将配体内吞过程中产生的力的关键作用与Notch结构研究结果结合起来。具体来说，我们假设配体结合的Notch的内吞作用产生强大的机械力，该机械力将Notch受体物理拉开以暴露ADAM切割位点并促进激活蛋白水解。我们的模型还提出，弗林蛋白酶裂解产生Notch异二聚体，使Notch易于通过配体内吞过程中产生的机械力发生物理解离。然而，配体内吞作用也被提议促进回收以产生高亲和力的Notch配体。为了解决这一争议，我们使用光镊来确定配体内吞作用在调节结合强度和力产生中的作用。 SMM 激光镊子用于测量 Notch 涂层珠子和配体表达细胞之间的内吞力和亲合力。寻找内吞力、粘附强度和Notch 胞吞作用之间的相关性。