New Amino Acids for Protein Engineering

用于蛋白质工程的新氨基酸

• 批准号: 6924753
• 负责人: DAVID A TIRRELL
• 金额: $ 34.17万
• 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 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6924753
• 关键词: alkynes; aminoacid analog; aminoacid tRNA ligase; aminoacid transport; azides; bioengineering /biomedical engineering; biomaterial development /preparation; biotechnology; crosslink; extracellular matrix proteins; high performance liquid chromatography; human tissue; isoleucine; ketones; leucine; matrix assisted laser desorption ionization; methionine; method development; molecular dynamics; nuclear magnetic resonance spectroscopy; phenylalanine analog; protein engineering; recombinant proteins; synthetic protein; tyrosine analog; valine; vascular endothelium

DESCRIPTION (provided by applicant): This application requests support for the development of new methods for the incorporation of novel amino acids into recombinant proteins. Computational and experimental studies will be combined to design mutant forms of the aminoacyl-tRNA synthetases that are capable of efficient activation of amino acid analogs bearing reactive functional groups. Special attention will be given to amino acids that carry alkyne, azide and ketone functional groups, in order to provide powerful new chemistries for use in the engineering of natural and artificial proteins. Computational design will be evaluated in comparison with experimental approaches that involve multiple rounds of mutagenesis and selection, and computational predictions will be assessed by comparison with the results of in vitro kinetic studies of amino acid activation. Cell free transcription - translation systems will be used as appropriate to identify and address limitations due to poor amino acid transport. The new chemistries to be developed in this work will open new opportunities in the design of engineered biomaterials.

描述（由申请人提供）：本申请请求支持开发将新型氨基酸掺入重组蛋白的新方法。计算和实验研究将结合起来，设计突变形式的氨酰-tRNA合成酶，能够有效激活的氨基酸类似物轴承反应性官能团。特别注意将给予携带炔，叠氮化物和酮官能团的氨基酸，以提供强大的新化学用于天然和人工蛋白质的工程。将通过与涉及多轮诱变和选择的实验方法进行比较来评估计算设计，并通过与氨基酸活化的体外动力学研究结果进行比较来评估计算预测。将酌情使用无细胞转录-翻译系统，以识别和解决由于氨基酸转运不良导致的限制。这项工作中开发的新化学物质将为工程生物材料的设计提供新的机会。