ROW: Expression and Sorting of the Myelin Protein Po

ROW：髓磷脂蛋白 Po 的表达和分选

• 批准号: 8720070
• 负责人: Marie Filbin
• 金额: $ 13.34万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1991-01-01
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8720070&HistoricalAwards=false
• 关键词: ROW; Expression; Sorting; Myelin; Protein

The formation of myelin by the Schwann cell requires the precise organization of a number of specific proteins. The long-term goals of this research are to understand how the Schwann cell delivers these proteins to the correct surface of the bipolar Schwann Cell. The immediate goals are to examine how the Schwann cell regulates the expression of one such protein, Po glycoprotein, when the external environment is changed. Because studies in vivo suggest that post-translational control of Po expression is important, the role of the sugar moieties in the expression of this glycoprotein will be investigated. To carry out these studies an abundance of Po is required. This will be accomplished by transfecting Schwann cells with the cDNA for Po followed by gene amplification. Furthermore, the transfected Po will be distinguished from the endogenously synthesized Po by attaching to the former an identifier/reporter peptide sequence. To ensure that Po with the reporter peptide is expressed/sorted in the same way as its endogenous counterpart, both will be characterized in transfected Chinese Hamster Ovary cells, prior to the studies with Schwann cells. Ultimately, the expression of Po-reporter will be monitored in Schwann cells in relation to the addition of axolemma and/or extracellular matrix to the culture before and after treatment specific inhibitors of glycosylation. Results from these studies can then be applied directly to Schwann cells in vivo and help elucidate the initial steps of myelinogenesis at the molecular level.

雪旺细胞形成髓鞘需要一系列特定蛋白质的精确组织。这项研究的长期目标是了解雪旺细胞如何将这些蛋白质运送到双极雪旺细胞的正确表面。目前的目标是研究当外部环境发生变化时，雪旺细胞如何调节一种这样的蛋白质--Po糖蛋白的表达。由于体内研究表明，翻译后对Po表达的控制是重要的，因此将研究糖部分在该糖蛋白表达中的作用。为了进行这些研究，需要大量的PO。这将通过将Po的cDNA导入雪旺细胞，然后进行基因扩增来完成。此外，通过在前者上附加一个识别符/报告肽序列，将转基因Po与内源性合成的Po区分开来。为了确保带有报告多肽的Po以与其内源性对应多肽相同的方式表达/分类，在用Schwann细胞进行研究之前，将在转基因的中国仓鼠卵巢细胞中对两者进行鉴定。最终，Po-Report在雪旺细胞中的表达将被监测与在处理特定的糖基化抑制剂之前和之后向培养物中添加轴膜和/或细胞外基质有关。这些研究的结果可以直接应用于体内的雪旺细胞，并有助于在分子水平上阐明髓鞘形成的初始步骤。