The chemical approach towards homogenous glycoprotein preparation and evaluation

制备均一糖蛋白的化学方法和 评估 摘要-我们的长期目标是发现细胞和感染性细胞的未知功能 通过询问单个均一的Pron糖型来获得Prion蛋白。这里的目标是 开发一类高效的利用应变分子的肽偶联方案，如β- 内酯和β-硫内酯，用于构建以前无法达到的目标，如环 四肽和膜蛋白。我们的中心假设，源自我们发表的 下面描述的结果和初步研究是，化学合成可以提供PrPC 和PrPSc菌株具有高构象保真度，这使得Prion生物学研究 在审问合成均一糖蛋白时，准确可靠地进行了检测。这个 修饰蛋白质结构的灵活性提供了以前无法获得的评估普里恩的工具 疾病。提出研究目标的理由是β-硫代内酯和β-内酯 释放环酯应变能够前所未有地快速构建酰胺键并连接 两个氨基酸残基。此外，环丁烯环的近平面几何构型将 通过防止恶唑酮的形成，生成所需的多肽加合物而不发生异构化。 一种类似的方法可以用来构建均一的膜糖蛋白 β-硫代内酯促进蛋白质连接。我们计划测试我们的中心假设，因此， 通过以下目标实现本应用程序的目标：1.设计和扩展 不需要异构化就可用于多肽合成的限制性酯的范围。2. 均一的细胞Pron糖蛋白及其同系物的合成。3.生物功能和 制备的多肽和蛋白质的活性研究。

The chemical approach towards homogenous glycoprotein preparation and evaluation ABSTRACT – Our long-term goal is to uncover the unknown function of cellular and infectious prion protein by interrogating individual homogenous prion glycoform. The objective here, is to develop a class of efficient peptidyl coupling protocols utilizing strained molecules, such as β- lactone and β-thiolactone, for the construction of previously unattainable targets such as cyclic tetrapeptides and membrane prion protein. Our central hypothesis, derived from our published results and preliminary studies described below, is that chemical synthesis can provide the PrPC and PrPSc strains with high conformational fidelity, which allows prion biological studies to be carried out with faithful accuracy when interrogating synthetic homogenous glyco-PrPs. The flexibility of modifying protein construct furnishes previously unavailable tool for evaluation prion disease. The rationale for the proposed research objectives is that the β-thiolactone and β-lactone release of cyclic ester strain enables unprecedented rapid amide bond construction and connects two amino acid residues. Furthermore, the near-planar geometry of the cyclobutene ring will generate desired peptidyl adducts without epimerization by preventing the oxazolone formation. An analogous approach could be applied to construct homogenous membrane glycoproteins from β-thiolactone promoted protein ligation. We plan to test our central hypothesis and, thereby, accomplish the objectives of this application via the following goals: 1. Design and expansion of the range of constrained esters that can be utilized for peptide synthesis without epimerization. 2. Synthesis of homogeneous cellular prion glycoproteins and congeners. 3. Biological function and activities investigation of prepared peptide and proteins.