Toolkit for Fast, Multipurpose and Inducible Bioorthogonal Chemistry

项目摘要 该提案的目标是开发一套多样化的新化学工具， 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正交试剂。将制定方法， 安全合成和直接偶联的'最低限度'四嗪的任何分子 感兴趣的，包括化学探针和荧光报告。我们亦建议 开发可用作“亲和生物正交化学”标签的四嗪， 在使蛋白质纯化和随后的位点选择性 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 大会提出。我们建议开发有效的催化方法， 在细胞环境中的快速生物正交化学，提供工具分子， 在细胞环境中的稳定性在其“关闭”状态，和最快的生物正交 反应在他们的“上”的状态。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开启。近ir 将开发光催化剂和热催化剂， 研究蛋白质动力学和蛋白质组学靶点鉴定的技术。 最后，我们还将开发生物正交N-酰基胞壁酸（NAM）和N- 用于微生物学中探针应用的酰基葡糖胺（NAG）分子， 免疫学研究。

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.