CAREER: Probing Structure and Expanding Function with Genetically Encoded Non- Canonical Amino Acids

职业：用基因编码的非规范氨基酸探索结构和扩展功能

• 批准号: 1847937
• 负责人: Christine Phillips-Piro
• 金额: $ 51.5万
• 依托单位: Franklin and Marshall College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1847937&HistoricalAwards=false
• 关键词: CAREER; Probing; Structure; Expanding; Function

Twenty amino acids naturally occur in living systems where they serve as building blocks of proteins. These proteins, in turn, have key functions in cells. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Christine Phillips-Piro from Franklin and Marshall College to use non-canonical amino acids (ncAAs), i.e., amino acids that do not occur in living systems, to investigate the structure and to systematically modify the function of proteins. The addition of new ncAAs results in unusual proteins that have very different functions than those made with the natural amino acids. The research project provides interdisciplinary training for undergraduate researchers in chemistry, molecular biology, biochemistry, and protein structure determination. Dr. Phillips-Piro's courses enable undergraduate students to learns about current scientific research. She integrates scientific literacy in two different courses - general chemistry and a general education course. Her team also helps to organize and run a regional chemistry conference for undergraduates (the Intercollegiate Student Chemists? Convention (ISCC)). Professor Phillips-Piro is a member of the Advanced Photon Source User Organization (APSUO) which seeks to broaden participation at Argonne National Laboratory. The research supported by this award exploits a targeted collection of ncAAs being used to both study protein structure and tune protein function. Local environments in a thermostable heme nitric oxide and/or oxygen binding protein (H-NOX) are assessed by using the ncAA vibrational reporter 4-cyano-L-phenylalanine in concert with temperature-dependent infrared spectroscopy and X-ray crystallography. The systematic modulation of the the oxygen binding affinity of the heme protein is achieved by using tyrosine ncAAs with altered phenolic hydrogen acidity. These moieties influence the strength of hydrogen bonding between a key amino acid residue (Y140) distal to the heme and the small molecule bound to the heme. Metal-binding ncAAs are used to generate a novel bimetallic enzyme based on the H-NOX scaffold. The insights gained and tools developed by Dr. Phillips-Piro may benefit other scientists who employ ncAAs to probe, modify, and/or engineer biological systems.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

二十种氨基酸自然出现在它们作为蛋白质基块的生活系统中。 这些蛋白质反过来在细胞中具有关键功能。有了这个奖项，化学生命过程的化学过程是为来自富兰克林和马歇尔学院的克里斯汀·菲利普斯·佩罗博士提供资金，用于使用非经典氨基酸（NCAAS），即生物中未发生的氨基酸，以研究结构并系统地修饰蛋白质的功能。添加新的NCAA会导致与天然氨基酸的功能截然不同的蛋白质。 该研究项目为化学，分子生物学，生物化学和蛋白质结构确定的本科研究人员提供了跨学科培训。 Phillips-Piro博士的课程使本科生能够了解当前的科学研究。她将科学素养整合到两个不同的课程中 - 通用化学和一项通识教育课程。她的团队还帮助组织和举办本科生的区域化学会议（大学生化学家？公约（ISCC））。 Phillips-Piro教授是高级光子源用户组织（APSUO）的成员，该组织旨在扩大在Argonne National Laboratory的参与。该奖项支持的研究利用了用于研究蛋白质结构和调整蛋白质功能的NCAA的靶向集合。通过与温度依赖性红外光谱光谱和X射线晶体学结合使用NCAA振动报告基因4-甲基-L-苯基丙氨酸，评估了可热稳定性血红素一氧化氮和/或氧结合蛋白（H-NOX）中的局部环境。血红素蛋白的氧结合亲和力的系统调节是通过使用酚类NCAA和酚酸酸度改变的。 这些部分会影响血红素远端的关键氨基酸残基（Y140）与与血红素结合的小分子之间的氢键强度。金属结合NCAA用于基于H-Nox支架产生一种新型的双金属酶。 Phillips-Piro博士获得的见解和工具可能会使其他使用NCAAS来探究，修改和/或工程师生物系统的科学家受益。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛的影响审查标准来评估值得通过评估的。