NEW AMINO ACIDS FOR PROTEIN ENGINEERING

用于蛋白质工程的新氨基酸

• 批准号: 6490168
• 负责人: DAVID A TIRRELL
• 金额: $ 29.62万
• 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/6490168
• 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：蛋白质工程是一个强大的工具，设计，如果新的液体 结晶相、大分子表面阵列、可逆水凝胶，以及 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制备 这些非天然蛋白质的高产量。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， 潜在的单体限于天然蛋白质氨基酸， 这些类似物可以被激活并带电荷来转移RNA。蒂瑞尔和 其他人已经成功地将蛋氨酸，异亮氨酸， 亮氨酸和苯丙氨酸通过它们各自的作用 氨酰-tRNA脱氢酶。具有烯属和炔属官能团的类似物 已经显示，在细菌蛋白质中， 合成.这些功能组的合并产生了重要的新的 化学衍生的机会，扩大材料的范围 这些特性可以设计成蛋白质聚合物。比如说， 烯烃复分解化学的最新进展导致 催化高效环化的过渡金属卡宾的开发 含有烯属侧链的肽。这项建议的目的是 结合联合收割机快速计算模拟筛选方法和实验-无论是在 体内和体外-寻找新的氨基酸用于蛋白质工程。 这种合作将导致快速和有效地发现非自然的 具有新的和有用的功能的氨基酸类似物，并将提供基础 用于构建具有所需性质的新型蛋白质样聚合物。的 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和瓦尔tRNA合成酶。