Toolkit for Fast, Multipurpose and Inducible Bioorthogonal Chemistry

项目摘要 这项提议的目标将是开发一套多样化的新化学工具 关于四嗪连接--已知最快的生物正交反应。这份提案描述了 继续努力开发新的生物正交试剂。将制定方法，用于 “极简主义”四嗪类化合物的安全合成及其与任何分子的直接偶联 兴趣，包括化学探针和荧光记者。我们还建议 开发可用作“亲和力生物正交化学”标签的四嗪类药物 在实现蛋白质纯化和随后的位点选择性方面起到双重作用 生物正交化学。在树脂蛋白质和蛋白质生物分子中的应用 提出了组件的概念。我们建议开发有效的催化方法来转化 在细胞环境中的快速生物正交化学，为工具分子提供高 处于关闭状态的细胞环境的稳定性，以及最快的生物正交性 到目前为止，人们对约会的反应都是“开”的。高效的光催化剂已经被开发出来 可以在体内和亚细胞水平上靶向生物正交开启。近红外 将开发光催化剂和热催化剂以及新的使能 蛋白质动力学研究和蛋白质组学靶标识别技术。 最后，我们还将开发生物正交N-酰基壁氨酸(NAM)和N- 酰氨基葡萄糖(NAG)分子，用于微生物学和生物化学中的探针应用 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.