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

• 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
• 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

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.

项目摘要 该提案的目的是开发一组以中心为中心的新化学工具 关于四嗪连接 - 已知最快的生物正交反应。该提案描述了 继续努力开发新的生物刚力试剂。将开发方法 “极简主义”四嗪的安全合成和直接耦合到任何分子 兴趣，包括化学问题和荧光记者。我们还建议 开发可以用作“亲和力生物正交化学”标签的四嗪 在启用蛋白质纯化，然后进行后续现场选择性中发挥双重作用 生物正交化学。应用于果仁蛋白蛋白质蛋白质和蛋白质 - 双分子 提出了组装。我们建议开发有效的催化方法 在细胞环境下快速生物正交化学，提供较高的工具分子 在其“偏离”状态下的细胞环境中的稳定性，最快的生物串管 迄今为止，其“在on”状态下的反应。已经开发了有效的光催化剂 可以在体内和亚细胞水平靶向生物正交的转盘。近ir 将开发光催化剂以及热催化剂 研究蛋白质动力学和蛋白质组学靶标识别的技术。 最后，我们还将开发生物正交的N-酰胺酸（NAM）和N- 酰基葡萄糖（NAG）分子用于微生物学和探针应用 免疫学研究。