Developing synthetic chemical biology strategies for biochemical investigations and biomedical applications

项目摘要 自然发生的和实验室的蛋白质修饰都是重要的研究领域，因为蛋白质是 几乎是每一个生物过程的中心。一方面，蛋白质翻译后修饰(PTM)是 对有机体的正常运作是必不可少的，与许多疾病有关。这是非常有意义的 研究PTMS对开发新的治疗药物具有重要意义。另一方面，继续 非典型氨基酸(NCAA)突变的研究进展为操作提供了有力的工具 以及蛋白质功能的研究。此Mira应用程序寻求合并这两个方面的调查 研究领域。这个Mira项目的第一个一般领域集中在共同翻译修改 通过基因密码工程获得蛋白质。长期目标是探索NCAA的创新战略 通过重新编程遗传密码进行突变。我们直接和独特的重点是重新编程 带有四重密码子的密码子语言(Qcodons)，尽管三重无意义密码子是主要的 用于遗传密码扩展领域的。基于我们在tRNA工程方面的开创性努力，我们 提出了通过识别和实现来提高Qcodon解码效率的新方向 对信号进行重新编码。使用重新编码信号也可以显著缓解对 带有基于无义抑制的非正则氨基的内源终止密码子的不良通读 酸(NCAA)在活细胞研究中的诱变作用。我们还将应用Qcodon依赖和NCAA介导的 控制战略到HIV-1疫苗的发展，这代表了一个新的方向，这是第一个 我的团队证明了这一点。这一战略也将应用于生产针对其他病毒的疫苗。 未来的致病病毒或细菌。米拉项目的第二个主要领域是调查 蛋白质酪氨酸O-硫化(PTS)在哺乳动物细胞生物学中的作用我们最初的努力将集中在以下几个方面 趋化因子受体作为我们长期努力研究PTS对受体信号的影响的第一步 总体而言。此外，我们还寻求开发针对硫酸盐受体的治疗剂。这两个 创新项目有自己的目标和意义，但在我们的专业知识下是统一的 化学/合成生物学，并部分地相互联系。我的实验室的长期目标是 不仅开发创新和有意义的化学/合成生物学工具，而且还使用这些工具 深入了解生物医学过程，以开发新的治疗方法。

Project Summary Both naturally occurring and laboratory protein modifications are important fields of research since proteins are central to virtually every biological process. On one hand, protein post-translational modifications (PTMs) are essential for proper functioning of an organism and are associated with many diseases. It is of great significance to study PTMs in order to develop new therapeutic agents. On the other hand, continuing advances in noncanonical amino acid (ncAA) mutagenesis has provided powerful tools for the manipulation and study of protein function. This MIRA application seeks to merge investigations on both fronts of these research areas. The first general area of this MIRA project focuses on the co-translational modification of protein through genetic code engineering. The long-term goal is to explore innovative strategies for ncAA mutagenesis through reprogramming the genetic code. Our immediate and unique focus is on reprogramming codon language with quadruplet codons (Qcodons), although triplet nonsense codons are the predominant ones used in the field of genetic code expansion. Built on our pioneering efforts on tRNA engineering, we propose a new direction to improve Qcodon decoding efficiency through the identification and implementation of recoding signals. The use of recoding signals can also significantly mitigate a major concern over undesirable readthrough of endogenous stop codons with nonsense suppression-based noncanonical amino acid (ncAA) mutagenesis in live cell studies. We will also apply a Qcodon-dependent and ncAA-mediated control strategy to the development of HIV-1 vaccines, which represents a novel direction that was first demonstrated by my group. Such strategy will also be applied to the generation of vaccines against other pathogenic viruses or bacteria in the future. The second general area of this MIRA project is to investigate the role of protein tyrosine O-sulfation (PTS) in mammalian cell biology. Our initial efforts will focus on PTS of chemokine receptors as the first step of our long-term efforts to study the effects of PTS on receptor signaling in general. Furthermore, we seek to develop therapeutic agents targeting sulfated receptors. These two innovative projects have their own goals and significance, but are unified under our expertise in chemical/synthetic biology and are partially associated with each other. The long-term goal of my laboratory is not only to develop innovative and meaningful chemical/synthetic biology tools, but also to employ these tools to gain insights into biomedical processes for the development of novel therapeutics.