Genetically Controlled Syntheses of New Polymeric Materials

新型高分子材料的基因控制合成

• 批准号: 9510031
• 负责人: David Tirrell
• 金额: $ 75.5万
• 依托单位: University of Massachusetts Amherst
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 1998-11-03
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9510031&HistoricalAwards=false
• 关键词: Genetically; Controlled; Syntheses; New; Polymeric

9510031 Tirrell This project will develop methods for the preparation of architecturally well-defined artificial proteins carrying reactive olefinic, dienyl, acetylenic and pyrrolyl functional groups. A key aspect of the project is the use of artificial genes to direct the synthesis of uniform polymer chains, which will acquire reactive functionality either directly (through the incorporation of unnatural amino acids), or indirectly (through post-translational modification reactions). In either approach, the fidelity of bacterial protein synthesis will be exploited to produce polymeric materials characterized by precise control of chain length, sequence, and stereochemistry. The direct approach will explore the translational activity of six artificial amino acids (and several variants thereof) related to isoleucine, phenylalanine, histidine or tyrosine. There is increasing evidence that the bacterial protein synthesis machinery can utilize a braoder range of amino acids than the twenty that are normally encoded by messenger RNA templates. The amino acid analogues chosen for this study are of particular interest owing to two facts: i). none of the natural amino acids carries olefinic, dienyl, acetylenic or pyrrolyl groups, and ii). such functional groups can be used to endow polymeric materials with interesting and potentially useful chemical and electrochemical properties. The approach to be used for direct incorporation involves: i). synthesis of an artificial gene encoding the sequence of interest, with the natural analogue encoded in place of the target unnatural amino acid, ii). cloning of the artificial gene in an appropriate auxotrophic bacterial strain, and iii). induction of target protein synthesis in a medium containing the unnatural amino acid (and generally depleted of the natural analogue). This strategy has been demonstrated successfully with several unnatural amino acids, including selenomethionine, p-fluorophenylalanine, 3- thienylalanine, trifluoroleucine, azetidinecarboxylic acid, dehydroproline, and thiaproline. Complementary strategy involving post-translational modification will also be explored. In this approach, protein biosynthesis will be used to prepare appropriately designed "base polymers," which will be modified with reagents carrying olefinic, dienyl, acetylenic or pyrrolyl groups. Polymers prepared by either route will be subjected to careful analyses of molecular structure, supramolecular organization, and functional properties. Particular attention will be given to the prospects for creating electrochemically switchable protein films containing pyrrolyl side chains. Such films would be of potential practical use as components of sensors, actuators and controlled delivery devices. %%% This research aims to carry out the synthesis of amino acid polymers with highly controlled structures, through the use of artificial genes. The polymers selected do not occur in nature and will be designed to have interesting electrical and other properties. ***

9510031 Tirrell该项目将开发制备结构明确的人工蛋白质的方法， 烯属的， 二烯基， 炔属 和 吡咯基官能团。 该项目的一个关键方面是使用人工基因来指导统一聚合物链的合成，这些聚合物链将直接（通过掺入非天然氨基酸）或间接（通过翻译后修饰反应）获得反应性功能。 在这两种方法中，细菌蛋白质合成的保真度将被利用来生产以精确控制链长、序列和立体化学为特征的聚合物材料。 直接方法将探索六种人工氨基酸（和几种变体）的翻译活性 其）与异亮氨酸、苯丙氨酸、组氨酸或酪氨酸相关。 越来越多的证据表明，细菌蛋白质合成机器可以利用比20种氨基酸更广泛的氨基酸， 通常都被编码 通过信使RNA模板。本研究选择的氨基酸类似物由于两个事实而特别令人感兴趣：i）。 天然氨基酸均不带有烯基、二烯基、炔基或吡咯基，和ii）。 这些官能团可用于赋予聚合物材料 有趣且潜在有用的化学物质 和电化学性质。 直接成立公司所采用的方法包括：i）. 合成编码感兴趣序列的人工基因，其中编码的天然类似物代替靶非天然氨基酸，ii）.在适当的营养缺陷型细菌菌株中克隆所述人工基因，和iii）. 在含有非天然氨基酸的培养基中诱导靶蛋白质合成（并且通常耗尽 的 天然类似物）。 这一战略 具有 已经成功地证明了几个非自然的 氨基酸，包括硒代蛋氨酸、对氟苯丙氨酸、3-噻吩丙氨酸、三氟亮氨酸、氮杂环丁烷羧酸、脱氢脯氨酸和硫代脯氨酸。 涉及翻译后修饰的补充策略也将 被探索。 在这种方法中，蛋白质生物合成将用于制备适当设计的“基础聚合物”，其将用携带烯属、二烯基、炔属或吡咯基的试剂进行修饰。 通过任何一种途径制备的聚合物都将经过分子量的仔细分析。 结构，超分子 组织， 和功能特性。 特别注意将给予电化学可切换的蛋白质薄膜含有吡咯侧链的前景。 这种薄膜作为传感器、致动器和受控释放装置的组件具有潜在的实际用途。 本研究的目的是通过使用人工基因来合成具有高度可控结构的氨基酸聚合物。 所选择的聚合物不存在于自然界中，并将被设计为具有有趣的电气和其他性能。 ***