Toolkit for Fast, Multipurpose and Inducible Bioorthogonal Chemistry

快速、多用途和诱导生物正交化学工具包

基本信息

批准号：
10660115
负责人：
JOSEPH M FOX
金额：
$ 49.35万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660115
关键词：
Acids Affinity Bacteria Binding Binding Proteins Biological Biological Response Modifier Therapy Biology Biotechnology Boronic Acids C-terminal Carbohydrates Cells Chemicals Chemistry Collaborations Coupling Cyclooctenes DNA Development Environment Family Generations Glycobiology Glycoconjugates Goals Heating Immunology Industrialization Label Ligands Ligation Light Methods Microbiology Modification N-terminal Nitrogen Organelles Organism Oxidation-Reduction Permeability Pharmacologic Substance Plant Resins Positioning Attribute Predisposition Preparation Procedures Property Protein Dynamics Protein Engineering Proteins Proteomics Radiochemistry Reaction Reagent Reporter Research Rhodamine Role Safety Series Side Site Techniques Technology Volatilization Work analog catalyst chelation commercialization experimental study fluorophore improved in vivo interest materials science metabolic engineering new technology novel protein purification scale up small molecule spatiotemporal tool two photon microscopy

项目摘要

Project Summary The goal of this proposal will be to develop a diverse set of new chemical tools centered on tetrazine ligation– the fastest known bioorthogonal reaction. This proposal describes a continued effort to develop new bioorthogonal reagents. Methods will be developed for the safe synthesis and direct coupling of ‘minimalist’ tetrazines to any molecule of interest, including chemical probes and fluorescent reporters. We also propose to develop tetrazines that can serve as “affinity bioorthogonal chemistry” tags that can serve a dual role in enabling protein purification followed by subsequent site-selective bioorthogonal chemistry. Applications to on-resin protein-protein and protein-biomolecule assembly are proposed. We propose to develop efficient catalytic methods for ‘turning on’ rapid bioorthogonal chemistry in cellular context, providing tool molecules with high stability in the cellular environment in their ‘off’ state, and the fastest bioorthogonal reactions to date in their ‘on’ state. Efficient photocatalysts have been developed that can target bioorthogonal turn-on in vivo and at the subcellular level. Near-IR photocatalysts as well as thermal catalysts will be developed as will new enabling technologies for studying protein dynamics and for proteomic target identification. Finally, we will also develop bioorthogonal N-acylmuramic acid (NAM) and N- acylglucosamine (NAG) molecules for use in probe applications in microbiology and immunology research.

项目概要该提案的目标是开发一套以关于四嗪连接——已知最快的生物正交反应该提案描述了。将继续努力开发新的生物正交试剂。 “极简”四嗪与任何分子的安全合成和直接偶联兴趣，包括化学探针和荧光生产者。开发可作为“亲和生物正交化学”标签的四嗪在实现蛋白质纯化和随后的位点选择性方面发挥双重作用生物正交化学在树脂上蛋白质-蛋白质和蛋白质-生物分子中的应用。我们建议开发有效的催化方法来进行“车削”。 on' 细胞环境中的快速生物正交化学，为工具分子提供了高细胞环境在“关闭”状态下的稳定性，以及最快的生物正交迄今为止，已开发出处于“开启”状态的高效光催化剂。可以在体内和亚细胞水平上靶向生物正交开启。将开发光催化剂和热催化剂以及新的使能技术研究蛋白质动力学和蛋白质组目标鉴定的技术。最后，我们还将开发生物正交的N-酰基胞壁酸（NAM）和N- 酰基葡萄糖胺 (NAG) 分子，用于微生物学和微生物学中的探针应用免疫学研究。