The chemical approach towards homogenous glycoprotein preparation and evaluation

均质糖蛋白制备和评价的化学方法

基本信息

批准号：
10654736
负责人：
Qiang Zhang
金额：
$ 36.56万
依托单位：
STATE UNIVERSITY OF NEW YORK AT ALBANY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

项目摘要

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.

制备均质糖蛋白的化学方法评估摘要 – 我们的长期目标是揭示细胞和感染性病毒的未知功能通过询问单个同质朊病毒糖型来检测朊病毒蛋白。利用应变分子（例如 β-）开发一类有效的肽基偶联方案内酯和 β-硫代内酯，用于构建以前无法实现的目标，例如环我们的中心假设源自我们已发表的文章。结果和初步研究如下所述，是化学合成可以提供PrPC 和具有高构象保真度的 PrPSc 菌株，这使得朊病毒生物学研究能够在询问合成均质糖-PrP 时，可以非常准确地进行。修饰蛋白质结构的灵活性为评估朊病毒提供了以前无法使用的工具所提出的研究目标的基本原理是β-硫内酯和β-内酯。环酯应变的释放实现了前所未有的快速酰胺键构建和连接两个氨基酸残基。此外，环丁烯环的近平面几何形状将通过防止恶唑酮形成，生成所需的肽基加合物，而不会发生差向异构化。类似的方法可以用于构建同质膜糖蛋白我们计划测试我们的中心假设，从而，通过以下目标实现本应用程序的目标： 1. 设计和扩展可用于肽合成而无需差向异构化的一系列受限酯。均质细胞朊病毒糖蛋白和同系物的合成 3. 生物学功能和制备的肽和蛋白质的活性研究。