Repurposing the Ribosome for Exotic Polymers

将核糖体重新用于外来聚合物

• 批准号: 9999711
• 负责人: Alanna Schepartz
• 金额: $ 6.67万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9999711
• 关键词: Repurposing; Ribosome; Exotic; Polymers

Project Summary: The machine used for template-guided protein synthesis–the ribosome and its associated translation factors–has been exploited for over 20 years to site-specifically incorporate > 150 unnatural α-amino acids and > 10 α-hydroxy acids into proteins, in vitro, in cells, and in animals. More exotic monomers, including N-alkyl α-amino acids, D-amino acids, and some β-amino acids have been incorporated using cell-free in vitro systems. These efforts have deepened our understanding of protein and RNA function and provided real-world commodities such as antibody-drug conjugates, modified therapeutics, and biomaterials to benefit society. Yet, even after two-plus decades of research, until very recently–the preliminary results for this application–there were no reports of a ribosome able to introduce a β-amino acid into a protein in vivo, nor any sophisticated bioinformatics tools for ribosome redesign. The goal of this application is to develop and apply genetic engineering, bioinformatics, and chemical biology tools to repurpose the bacterial translational apparatus for the templated, in vivo biosynthesis of proteins and polypeptides containing backbone-modified monomers. The primary focus is on β- amino acids, as preliminary results indicate that this goal is attainable with a 4-5 year RO1 award, but the strategies, insights, and tools developed will provide a robust infrastructure for the sequence-templated biosynthesis of diverse classes of evolvable matter–truly exotic biopolymers–whose functions are limited only by our collective imagination. By creating new machines that efficiently incorporate β-amino acids into proteins in vivo, we will not only provide tools to address enduring questions about natural translation, but also provide real benefit to diverse segments of bio-medical research engine.

项目概要： 用于模板引导蛋白质合成的机器——核糖体及其相关物 翻译因素 - 已被利用 20 多年，以特定地点合并 > 150 在体外、细胞和动物体内将非天然 α-氨基酸和 > 10 种 α-羟基酸转化为蛋白质。 更多外来单体，包括 N-烷基 α-氨基酸、D-氨基酸和一些 β-氨基 已使用无细胞体外系统掺入酸。这些努力加深了我们 了解蛋白质和 RNA 功能并提供现实世界的商品，例如 抗体药物偶联物、改良疗法和生物材料造福社会。然而，即使 经过两年多的研究，直到最近——初步结果 应用——没有关于核糖体能够将β-氨基酸引入蛋白质的报道 体内，也没有任何用于核糖体重新设计的复杂生物信息学工具。此举的目标 应用是基因工程、生物信息学、化学生物学的开发和应用 重新利用细菌翻译装置进行模板化体内生物合成的工具 含有骨架修饰单体的蛋白质和多肽。主要焦点是β- 氨基酸，初步结果表明这一目标可以通过 4-5 年的 RO1 实现 奖，但开发的策略、见解和工具将为 不同类别可进化物质的序列模板生物合成——真正的异国情调 生物聚合物——其功能仅受我们集体想象力的限制。通过创造新的 能够在体内有效地将 β-氨基酸整合到蛋白质中的机器，我们不仅提供 解决有关自然翻译的持久问题的工具，同时也为 生物医学研究引擎的各个细分领域。