Collaborative Research: Repurposing the translation apparatus for mirror image polypeptide synthesis

合作研究：重新利用镜像多肽合成的翻译装置

• 批准号: 1716766
• 负责人: Michael Jewett
• 金额: $ 47.5万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1716766&HistoricalAwards=false
• 关键词: Collaborative; Research; Repurposing; translation; apparatus

The translation apparatus is the cell's factory for protein biosynthesis, stitching together amino acid substrates into sequence-defined polymers (proteins) from a defined genetic template. The extraordinary synthetic capability of the protein biosynthesis system has driven extensive efforts to harness it for societal needs in areas as diverse as energy, materials, and medicine. For example, recombinant protein production has transformed the lives of millions of people through the synthesis of biopharmaceuticals and industrial enzymes. In nature, however, only limited sets of protein monomers are utilized, thereby resulting in limited sets of biopolymers (i.e., proteins). Expanding nature's repertoire of ribosomal monomers could yield new classes of enzymes, therapeutics, materials, and chemicals with diverse chemistry. In the short term, this will expand the genetic code in a unique and transformative way. In the long-term, knowledge gained will allow researchers to diversify, evolve and repurpose the ribosome and the entire protein synthesis system to generate non-natural polymers as new classes of sequence-defined, evolvable matter. This proposal will also promote interdisciplinary education, including the specific expansion of STEM education and career opportunities for underrepresented minorities and women. Students will be trained to integrate principles from genome engineering, systems biology, and synthetic biology. As a form of outreach, the investigators will create experiential learning modules that bring synthetic biology research to K-12 and undergraduate classrooms and connect students to the science being done at our institutions. This new outreach program will ensure that advances made in this project benefit a broader community and will contribute to motivating and training young scientists and engineers. In this project, the investigators seek to repurpose the translation apparatus for making new proteins containing multiple mirror-image D-alpha-amino acids. By seamlessly melding the integration of bottom-up engineering design, genome engineering, and full-scale systems optimization, the project will create a new framework for studying and engineering translation and the genetic code. This framework will be instrumental in sustaining the much-anticipated transformation in expanding the range of genetically encoded chemistry in living systems, with the potential for significant breakthroughs. For example, understanding the structural and substrate flexibility of the protein synthesis machinery may provide insights into life's origin and also lead to general rules for engineering protein synthesis to meet societal needs. From an engineering perspective, the research could enable scalable mirror image polypeptide synthesis, opening the door to cheaper and more effective peptidomimetic enzymes, materials, and drugs, and expanding the scope of synthetic and chemical biology. In sum, it is expected that this project will provide new directions for academic and industrial enterprises related to engineering translation, while simultaneously training the next generation of scientists and engineers to be full participants in the work force.

翻译装置是细胞蛋白质生物合成的工厂，将氨基酸底物从定义的遗传模板拼接成序列定义的聚合物（蛋白质）。蛋白质生物合成系统非凡的合成能力推动了广泛的努力，以利用它来满足能源，材料和医学等领域的社会需求。例如，重组蛋白质生产通过生物制药和工业酶的合成改变了数百万人的生活。然而，在自然界中，仅利用有限组的蛋白质单体，从而导致有限组的生物聚合物（即，蛋白质）。扩大自然界的核糖体单体库可以产生新类别的酶，治疗剂，材料和具有不同化学性质的化学品。在短期内，这将以一种独特和变革性的方式扩展遗传密码。从长远来看，所获得的知识将使研究人员能够使核糖体和整个蛋白质合成系统多样化，进化和重新定位，以产生非天然聚合物作为新的序列定义的可进化物质。该提案还将促进跨学科教育，包括为代表性不足的少数民族和妇女具体扩大STEM教育和职业机会。学生将接受培训，以整合基因组工程，系统生物学和合成生物学的原则。作为一种推广形式，研究人员将创建体验式学习模块，将合成生物学研究带到K-12和本科课堂，并将学生与我们机构正在进行的科学联系起来。这项新的推广计划将确保该项目取得的进展使更广泛的社区受益，并将有助于激励和培训年轻的科学家和工程师。在这个项目中，研究人员试图重新利用翻译装置来制造含有多个镜像D-α-氨基酸的新蛋白质。通过无缝融合自下而上的工程设计、基因组工程和全面系统优化，该项目将为研究和工程翻译以及遗传密码创建一个新的框架。这一框架将有助于维持人们期待已久的转变，扩大生命系统中遗传编码化学的范围，并有可能取得重大突破。例如，了解蛋白质合成机制的结构和底物灵活性可能会提供对生命起源的见解，并导致工程蛋白质合成的一般规则，以满足社会需求。从工程角度来看，这项研究可以实现可扩展的镜像多肽合成，为更便宜，更有效的肽模拟酶，材料和药物打开大门，并扩大合成和化学生物学的范围。总之，预计该项目将为与工程翻译相关的学术和工业企业提供新的方向，同时培训下一代科学家和工程师成为劳动力的全面参与者。

项目成果

A Highly Productive, One-Pot Cell-Free Protein Synthesis Platform Based on Genomically Recoded Escherichia coli

• DOI: 10.1016/j.chembiol.2019.10.008
• 发表时间: 2019-12-19
• 期刊: CELL CHEMICAL BIOLOGY
• 影响因子: 8.6
• 作者: Des Soye, Benjamin J.;Gerbasi, Vincent R.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.

Repurposing ribosomes for synthetic biology.

• DOI: 10.1016/j.cbpa.2017.07.012
• 发表时间: 2017-10
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8
• 作者: Liu Y;Kim DS;Jewett MC
• 通讯作者: Jewett MC

Engineered ribosomes with tethered subunits for expanding biological function

• DOI: 10.1038/s41467-019-11427-y
• 发表时间: 2019-09-02
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Carlson, Erik D.;D'Aquino, Anne E.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.

Next-generation genetic code expansion.

• DOI: 10.1016/j.cbpa.2018.07.020
• 发表时间: 2018-10
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8
• 作者: Arranz-Gibert P;Vanderschuren K;Isaacs FJ
• 通讯作者: Isaacs FJ

In vitro ribosome synthesis and evolution through ribosome display

• DOI: 10.1038/s41467-020-14705-2
• 发表时间: 2020-02-28
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Hammerling, Michael J.;Fritz, Brian R.;Jewett, Michael C.
• 通讯作者: Jewett, Michael C.