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: 1714860
• 负责人: Farren Isaacs
• 金额: $ 47.5万
• 依托单位: Yale 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 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1714860&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.

翻译装置是细胞的蛋白质生物合成工厂，将氨基酸底物缝合在一起，根据定义的遗传模板形成序列定义的聚合物(蛋白质)。蛋白质生物合成系统的非凡合成能力推动了广泛的努力，以利用它来满足能源、材料和医药等各种领域的社会需求。例如，重组蛋白生产通过合成生物制药和工业酶改变了数百万人的生活。然而，在自然界中，只有有限的蛋白质单体被利用，从而导致生物聚合物(即蛋白质)的有限集合。扩展自然界的核糖体单体系列可能会产生具有不同化学成分的新类别的酶、治疗药物、材料和化学品。在短期内，这将以一种独特和变革性的方式扩大遗传密码。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类别的序列定义的可进化物质。这项提案还将促进跨学科教育，包括具体扩大STEM教育，并为代表不足的少数群体和妇女提供就业机会。学生将接受综合基因组工程、系统生物学和合成生物学原理的培训。作为一种外展形式，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。这一新的推广计划将确保该项目取得的进展惠及更广泛的社区，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，研究人员试图改变翻译装置的用途，以制造包含多个镜像D-α-氨基酸的新蛋白质。通过将自下而上的工程设计、基因组工程和全面的系统优化无缝融合，该项目将创建一个研究和工程翻译以及遗传密码的新框架。这一框架将有助于维持备受期待的转变，扩大生物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的灵活性可能会提供对生命起源的洞察，也会导致工程蛋白质合成以满足社会需求的一般规则。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、更有效的多肽仿酶、材料和药物打开大门，并扩大合成和化学生物学的范围。综上所述，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，同时培养下一代科学家和工程师成为工作的全面参与者。

项目成果

Encoding human serine phosphopeptides in bacteria for proteome-wide identification of phosphorylation-dependent interactions.

• DOI: 10.1038/nbt.4150
• 发表时间: 2018-08
• 期刊: Nature biotechnology
• 影响因子: 46.9
• 作者: Barber KW;Muir P;Szeligowski RV;Rogulina S;Gerstein M;Sampson JR;Isaacs FJ;Rinehart J
• 通讯作者: Rinehart J

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