Role of E-cadherin’s N-glycans in tissue morphogenesis

E-钙粘蛋白的 N-聚糖在组织形态发生中的作用

• 批准号: 525776139
• 负责人: Professor Dr. Jörg Großhans
• 金额: --
• 依托单位: Arbeitsgruppe Entwicklungsgenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/525776139?language=en
• 关键词: Role; cadherin; glycans; tissue; morphogenesis

The evolutionary conserved E-cadherin (shotgun in Drosophila) is the prototype of a cell adhesion molecule in epithelia, which on the one side is responsible for homotypic and Calcium dependent cell-cell adhesion and on the other is connected to the cortical actomyosin cytoskeleton by forming a trimeric complex with beta-catenin (armadillo in Drosophila) and alpha-catenin. Beside such generic trimeric complexes, E-cadherin is found in super-molecular complexes of several hundred molecule, which may or may not be engaged in adhesion and trimeric complexes. Being centrally positioned at the force transmission between epithelial cells, E-cadherin plays a pivotal role in mechanical cell communication. The molecular mechanism of mechanotransduction has been difficult to investigate given the intertwined E-cadherin functions in adhesion, cortical link and mechanotransduction. Based on findings that hypo-N-glycosylated E-cadherin is impaired in clustering, cortical interactions and certain aspects of cell-cell communication, we propose to systematically investigate the contributions of N-glycans to the functions of E-cadherin in a physiological setting during tissue morphogenesis in Drosophila embryos by a series of E-cadherin mutations at N-glycosylation sites. The phenotype of these mutations will be investigated by established molecular and functional assays for E-cadherin mobility, complex formation, mechanics of junctions, adhesion, cell signalling. The phenotypes in nano-structure will be investigated by high resolution microscopy and DNA-paint single molecular localization.

进化上保守的E-钙粘蛋白（果蝇中的鸟枪蛋白）是上皮细胞中细胞粘附分子的原型，其一方面负责同型和钙依赖性细胞-细胞粘附，另一方面通过与β-连环蛋白（果蝇中的犰狳蛋白）和α-连环蛋白形成三聚体复合物而连接至皮质肌动球蛋白细胞骨架。除了这些一般的三聚体复合物之外，在数百个分子的超分子复合物中发现了E-钙粘蛋白，其可以参与或不参与粘附和三聚体复合物。E-cadherin位于上皮细胞之间的力传递中心，在细胞机械通讯中起着关键作用。由于E-cadherin在粘附、皮质连接和机械传导中的功能交织在一起，因此机械传导的分子机制一直难以研究。基于低N-糖基化的E-钙粘蛋白在聚类、皮质相互作用和细胞-细胞通讯的某些方面受损的研究结果，我们提出系统地研究N-聚糖在果蝇胚胎组织形态发生过程中对E-钙粘蛋白功能的贡献，通过一系列N-糖基化位点的E-钙粘蛋白突变。这些突变的表型将通过已建立的E-钙粘蛋白迁移率、复合物形成、连接力学、粘附、细胞信号传导的分子和功能测定进行研究。纳米结构中的表型将通过高分辨率显微镜和DNA-paint单分子定位进行研究。