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

20种氨基酸自然存在于生命系统中，它们是蛋白质的组成部分。反过来，这些蛋白质在细胞中具有关键功能。化学部的生命过程化学项目将资助富兰克林和马歇尔学院的Christine Phillips-Piro博士使用非规范氨基酸(NCAA)，即生命系统中不存在的氨基酸，来研究蛋白质的结构和系统地修饰蛋白质的功能。新的NCAA的加入导致了不寻常的蛋白质，其功能与用天然氨基酸制成的蛋白质非常不同。该研究项目为本科生研究人员提供化学、分子生物学、生物化学和蛋白质结构测定方面的跨学科培训。菲利普斯-皮罗博士的课程使本科生能够了解当前的科学研究。她将科学素养融入了两门不同的课程--普通化学和普通教育课程。她的团队还帮助组织和举办了一次面向本科生的地区性化学会议(校际学生化学家？《公约》))。菲利普斯-皮罗教授是高级光子源用户组织(APSUO)的成员，该组织寻求扩大阿贡国家实验室的参与。这项由该奖项支持的研究利用了一组有针对性的NCAA，用于研究蛋白质结构和调节蛋白质功能。用NCAA振动报告4-氰基-L-苯丙氨酸结合变温红外光谱和X射线结晶学研究了耐热血红素、一氧化氮和/或氧结合蛋白(H-NOx)中的局部环境。系统地调节血红素蛋白的氧结合亲和力是通过使用具有改变的酚氢酸度的酪氨酸NCAA来实现的。这些部分会影响亚铁血红素远端的关键氨基酸残基(Y140)与与亚铁血红素结合的小分子之间的氢键强度。金属结合的NCAA被用来合成一种基于H-NOX支架的新型双金属酶。菲利普斯-皮罗博士获得的洞察力和开发的工具可能会让其他雇用NCAA来探测、修改和/或设计生物系统的科学家受益。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.