Mechanism-inspired development of a chemoselective click-based bio-orthogonal reaction platform

基于化学选择性点击的生物正交反应平台的机制启发开发

• 批准号: 2098230
• 金额: --
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2098230
• 关键词: Mechanism; inspired; development; chemoselective; click

Background - The post-genomic era has provided biologists with a wealth of information and -omic sequencing tools to further our understanding of fundamental biology and disease. However, it is still exceedingly difficult to interrogate biological processes where the link between gene and protein expression is decoupled. A pertinent exemplar of this is changes in the glycosylation state between healthy and cancerous cells presented on the cell surface. An emerging tool used to interrogate these changes is the metabolic incorporation of 'chemical reporters' within a target biomolecule and access to highly specific 'bio-orthogonal' chemical reactions, which preferably react only with the chemical reporter group. Key to the success of this approach is the availability of bio-orthogonal reactions that are fast, biocompatible, and chemoselective.The incorporation of two chemical reporters followed by chemoselective bio-orthogonal labelling is an extension of this 'tag and modify' strategy, which offers the opportunity to label two different biomolecules in cellulo. Additionally, the incorporation of different chemical reporters within a single biomolecule followed by dual differential labelling is a nascent approach to probe the dynamics of a single biomolecule. At present, this is highly challenging to achieve with current bio-orthogonal strategies if discretely tagged products (i.e., without a mixture of products) are required.Previous work - Our collaborative team has recently identified the utility of aromatic ynamines as a superior reagent for Cu-catalyzed alkyne-azide cycloaddition (CuAAC) reactions. These alkyne surrogates require significantly less Cu catalyst are the only alkyne reagents reported to date which enable chemoselective control in a sequential two-step CuAAC process.Project Objective - The principal objective of this studentship is to develop aromatic ynamines into a powerful new bio-orthogonal reaction platform for the chemoselective tagging of glycoproteins in prostate cancer cells. The specific aims of the project are to:(i) gain a mechanistic understanding of the enhanced chemoselectivity of ynamines in CuAAC reactions, and potentially across other bio-orthogonal reaction classes.(ii) establish conditions for chemoselective modification of biomolecules.(iii) explore the utility of ynamines as glycoprotein tagging agents of prostate cancer cells.

背景-后基因组时代为生物学家提供了丰富的信息和基因组测序工具，以促进我们对基础生物学和疾病的理解。然而，它仍然是非常困难的询问生物过程中的基因和蛋白质表达之间的联系是解耦的。一个相关的例子是细胞表面上健康细胞和癌细胞之间糖基化状态的变化。一种用于询问这些变化的新兴工具是在靶生物分子内代谢掺入“化学报告物”，并进入高度特异性的“生物正交”化学反应，其优选仅与化学报告基团反应。这种方法成功的关键是生物正交反应的可用性，是快速的，生物相容性，和chemoselictional.The纳入两个化学报告随后由chemoselictional-orthogonal标记是这种“标签和修改”策略的延伸，它提供了机会，以标记两个不同的生物分子在cellulo。此外，在单个生物分子内掺入不同的化学报告分子，然后进行双重差异标记是探测单个生物分子动力学的新生方法。目前，如果离散标记的产物（即，以前的工作-我们的合作团队最近已经确定了芳香族炔类化合物作为Cu催化的炔-叠氮化物环加成（CuAAC）反应的上级试剂的效用。这些炔替代品需要显着较少的铜催化剂是唯一的炔试剂报告日期，使化学选择性控制在一个连续的两步CuAAC process.Project目标-本研究的主要目标是开发芳香族化合物成为一个强大的新的生物正交反应平台的糖蛋白在前列腺癌细胞的化学选择性标签。该项目的具体目标是：（i）获得对CuAAC反应中，以及潜在的其他生物正交反应类别中，精氨酸的化学选择性增强的机械理解。(ii)建立生物分子化学选择性修饰的条件。(iii)探索将甜菜碱作为前列腺癌细胞的糖蛋白标记剂的效用。