New Amino Acids for Protein Engineering

描述(由申请人提供)：本申请请求支持将新的氨基酸加入重组蛋白质的新方法的开发。计算和实验研究将结合起来，设计能够有效激活带有反应性官能团的氨基酸类似物的氨基酰基-tRNA合成酶的突变形式。将特别关注携带炔基、叠氮基和酮基的氨基酸，以便为在天然和人造蛋白质工程中使用提供强大的新化学物质。计算设计将通过与涉及多轮突变和选择的实验方法进行比较来评估，计算预测将通过与氨基酸激活的体外动力学研究结果进行比较来评估。细胞自由转录-翻译系统将被适当地用来识别和解决由于氨基酸转运不良而造成的限制。将在这项工作中开发的新化学将为工程生物材料的设计带来新的机会。

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.