Developing synthetic chemical biology strategies for biochemical investigations and biomedical applications

项目摘要 天然存在的和实验室蛋白质修饰都是重要的研究领域，因为蛋白质是 对几乎所有生物过程都至关重要一方面，蛋白质翻译后修饰（PTM）是 对有机体的正常功能至关重要，并与许多疾病有关。具有重要 因此，研究PTM对开发新的治疗药物具有重要意义。另一方面，继续 非规范氨基酸（ncAA）诱变的进展为操作提供了强有力的工具 研究蛋白质的功能。这MIRA申请旨在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翻译修饰， 蛋白质的基因编码。长期目标是探索ncAA的创新策略 通过重新编程遗传密码进行诱变。我们当前和独特的重点是重新编程 密码子语言与四联体密码子（Qcodons），虽然三联体无义密码子是主要的 用于遗传密码扩展的基因基于我们在tRNA工程方面的开创性努力，我们 通过识别和实现，提出了提高Qcodon解码效率的新方向 重新编码信号。使用重新编码信号还可以显著减轻对以下问题的主要担忧： 具有基于无义抑制的非规范氨基酸的内源终止密码子的不希望的通读 酸（ncAA）诱变的研究。我们还将应用Q密码子依赖性和ncAA介导的 HIV-1疫苗的开发控制策略，这代表了一个新的方向， 由我的团队演示。这种策略也将应用于产生针对其他疾病的疫苗。 致病性病毒或细菌。这个MIRA项目的第二个一般领域是调查 蛋白质酪氨酸O-硫酸化（PTS）在哺乳动物细胞生物学中的作用。我们的初步工作将侧重于 作为我们长期研究PTS对受体信号传导影响的第一步， 梗概.此外，我们寻求开发靶向硫酸化受体的治疗剂。这两 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，并且彼此部分相关。我实验室的长期目标是 不仅要开发创新和有意义的化学/合成生物学工具，而且要使用这些工具， 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.