NEW AMINO ACIDS FOR PROTEIN ENGINEERING

用于蛋白质工程的新氨基酸

• 批准号: 6254772
• 负责人: DAVID A TIRRELL
• 金额: $ 29.52万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6254772
• 关键词: aminoacid; aminoacid analog; aminoacid tRNA ligase; aminoacyl tRNA; binding sites; bioengineering /biomedical engineering; biomaterial development /preparation; biotechnology; chemical group; chemical kinetics; chemical structure function; combinatorial chemistry; computer simulation; elastin; intermolecular interaction; isoleucine; methionine; molecular biology information system; molecular energy level; peptide chemical synthesis; peptide library; protein engineering; site directed mutagenesis; tissue engineering; transfection /expression vector

DESCRIPTION: Protein engineering is a powerful tool for design if novel liquid crystal phases, macromolecular surface arrays, reversible hydrogels, and artificial extracellular matrices for use in tissue regeneration and repair. In vivo microbial expression of artificial genes provides a means of preparing such non-natural proteins in high yields. The target structure is encoded into an artificial gene, and the gene is expressed in an appropriate microbial host. However, in vivo protein engineering poses a challenge in that the pool of potential monomers is restricted to the natural proteinogenic amino acids and those analogs that can be activated and charged to transfer RNAs. Tirrell and others have successfully incorporated analogs of methionine, isoleucine, leucine, and phenylalanine through the action of their respective aminoacyl-tRNA synthases. Analogs with olefinic and acetylenic functional groups have been shown to serve as methionine surrogates in bacterial protein synthesis. Incorporation of such functional groups creates important new opportunities for chemical derivatization, extending the range of materials properties that can be designed into protein-based polymers. For example, recent advances in the chemistry of olefin metathesis have led to the development of transition metal carbenes that catalyze efficient cyclization of peptides containing olefinic side chains. The objective of this proposal is to combine fast computational analog screening methods and experiments - both in vivo and in vitro - to find new amino acids for use in protein engineering. This collaboration will lead to fast and efficient discovery of non-natural amino acid analogs with new and useful functionality and will provide a basis for building novel protein-like polymers with desired properties. The computational methods to be used here have already been tested for design of analogs for phenylalanine and will be extended to new substrates for Phe, Met, Ile, Leu, and Val tRNA synthetases.

描述：蛋白质工程是新型液体设计的强大工具 晶相、大分子表面阵列、可逆水凝胶和 用于组织再生和修复的人工细胞外基质。在 人工基因的体内微生物表达提供了一种制备方法 这种非天然蛋白质的产量很高。目标结构被编码为 人工基因，并且该基因在适当的微生物宿主中表达。 然而，体内蛋白质工程提出了一个挑战，因为 潜在的单体仅限于天然蛋白质氨基酸和 那些可以被激活并带电以转移 RNA 的类似物。蒂雷尔和 其他人已成功掺入蛋氨酸、异亮氨酸的类似物， 亮氨酸和苯丙氨酸通过各自的作用 氨酰-tRNA 合酶。具有烯属和炔属官能团的类似物 组已被证明可作为细菌蛋白质中的蛋氨酸替代物 合成。这些功能组的结合创造了重要的新功能 化学衍生化的机会，扩大材料的范围 可以设计成基于蛋白质的聚合物的特性。例如， 烯烃复分解化学的最新进展导致 开发催化高效环化的过渡金属卡宾 含有烯属侧链的肽。该提案的目的是 结合快速计算模拟筛选方法和实验 - 都在 体内和体外 - 寻找用于蛋白质工程的新氨基酸。 此次合作将导致快速有效地发现非自然物质 具有新的和有用的功能的氨基酸类似物，并将提供基础 用于构建具有所需特性的新型类蛋白质聚合物。这 这里使用的计算方法已经过设计测试 苯丙氨酸的类似物，并将扩展到 Phe、Met、 Ile、Leu 和 Val tRNA 合成酶。