ROLE OF N-GLYCOSYLATION IN E-CADHERIN MEDIATED CELL-CELL ADHESION

N-糖基化在 E-钙粘蛋白介导的细胞粘附中的作用

• 批准号: 7602000
• 负责人: MARIA A. KUKURUZINSKA
• 金额: $ 3.88万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-03 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602000
• 关键词: Adherens Junction; Adhesions; Binding; Cadherins; Cell Communication; Cell-Cell Adhesion; Cells; Characteristics; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; E-Cadherin; Epithelial Cells; Event; Fingerprint; Funding; Gel; Grant; Institution; Investigation; Mediating; Membrane Glycoproteins; Membrane Proteins; Methodology; Methods; Modification; Molecular; Non-Malignant; Pattern; Peptides; Polysaccharides; Protein Binding; Proteins; Research; Research Personnel; Resources; Role; Signal Transduction; Site; Source; United States National Institutes of Health; Weight; Western Blotting; Work; cancer cell; cancer type; extracellular; glycosylation; insight; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. E-cadherin is a 120-kDa membrane glycoprotein, expressed in epithelial cells, which is the main player in establishing adherens junctions between cells. The adherens junctions have both intra and extracellular anchorage points in order to regulate cell-cell interactions.[1-2] Alterations in the assembly or disassembly of adherens junctions occur in association with major changes in the state of cell, including differentiation and proliferation [3], as well as in cancer progression. Loss of E-cadherin function is a frequent event in many types of cancer, commonly caused by diminished or aberrant E-cadherin expression. [4] Recent studies have indicated that the N-glycosylation pattern of E-cadherin has a role in the molecular organization of adherens junctions. Specifically, the presence of complex N-glycans is associated with destabilized adherens junctions. [5] Investigation into the structural characteristics of E-cadherin, specifically its N-glycosylation pattern and protein binding partners, provide insight into E-cadherin mediated adhesion and cell signaling functions. Isolation of this large membrane protein has required the development of specific methods. Our methodology has isolated pools of E-cadherin with different binding partners and different N-glycosylation states, depending on cell status. The forms of E-cadherin in malignant cells are more highly glycosylated than are those in non-malignant cells; they migrate at a higher apparent weight, as detected by Western blot and peptide mass fingerprinting of excised SDS-PAGE bands. Through continued studies to identify the protein components, we have found that E-cadherin is associated with destabilizing proteins in malignant cells to a greater extent than when it is present in non-malignant cells. E-cadherin N-glycosylation has also been investigated, specifically site occupancy and N-glycan composition. MALDI-TOF MS analysis of in-gel enzymatically released N-glycans suggests E-cadherin contains predominantly complex N-glycans. This work highlights that cell context, the recruitment of specific proteins to the adhesion complex, along with specific structural modifications to E-cadherin may define the overall stability of E-cadherin mediated adhesion. References: 1. Gottardi CJ, Wong E, Gumbiner BM. J Cell Biol 2001; 153(5): 1049-60 2. Perez-Moreno M, Jamora C, Fuchs E. Cell 2003; 112(4): 535-48 3. Gumbiner BM. Nat Rev Mol Cell Biol 2005; 6:622-634 4. Wheelock MJ, Johnson KR. Annu Rev Cell Dev Biol 2003; 19:207-35 5. Liwosz A, Lei T, and Kukuruzinska MA. J Biol Chem 2006; 281:23138-49.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 E-钙粘着蛋白是一种120 kDa膜糖蛋白，在上皮细胞中表达，这是在细胞之间建立粘附连接的主要参与者。粘附剂连接具有调节细胞 - 细胞相互作用的细胞内和细胞外锚固点。[1-2]粘附连接的组装或拆卸的改变发生在细胞状态的重大变化，包括分化和增殖[3]以及癌症进展。在许多类型的癌症中，E-钙粘蛋白功能的丧失是经常发生的事件，通常是由E-钙粘蛋白表达降低或异常引起的。 [4]最近的研究表明，e-钙粘蛋白的N-糖基化模式在粘附连接的分子组织中起作用。具体而言，复杂的N-聚糖的存在与不稳定的粘附连接有关。 [5]研究E-钙粘蛋白的结构特征，特别是其N-糖基化模式和蛋白结合伴侣，可深入了解E-钙粘着蛋白介导的粘附和细胞信号传导功能。 这种大膜蛋白的分离需要开发特定方法。 我们的方法具有不同的结合伴侣和不同的N-糖基化态的E-钙粘蛋白池，具体取决于细胞状态。 与非恶性细胞相比，恶性细胞中的E-钙粘蛋白的形式更高。它们以更高的明显重量迁移，如切除的SDS-PAGE带的Western印迹和肽质量指纹所检测到的。 通过持续的研究以鉴定蛋白质成分，我们发现E-钙粘着蛋白与在非恶性细胞中存在的恶性细胞中的蛋白质不稳定有关。 还研究了E-钙粘蛋白N-糖基化，特别是位点占用率和N-聚糖组成。 MALDI-TOF MS分析酶内释放的N-聚糖表明E-钙粘蛋白主要含有复杂的N-聚糖。 这项工作强调了细胞环境，特定蛋白质对粘附复合物的募集以及对电子钙粘着蛋白的特定结构修饰可能定义了电子钙粘着蛋白介导的粘附的整体稳定性。 参考： 1。GottardiCJ，Wong E，Gumbiner BM。 J Cell Biol 2001； 153（5）：1049-60 2。Perez-Moreno M，Jamora C，Fuchs E. Cell 2003; 112（4）：535-48 3。GumbinerBM。 Nat Rev Mol Cell Biol 2005; 6：622-634 4。WheelockMJ，Johnson Kr。 Annu Rev Cell Dev Biol 2003; 19：207-35 5。LiwoszA，Lei T和Kukuruzinska MA。 J Biol Chem 2006; 281：23138-49。