CAREER: Expanding Protein Chemistry through the Evolution of Orthogonal Non-Natural Heme:Enzyme Pairs

职业：通过正交非天然血红素：酶对的进化扩展蛋白质化学

• 批准号: 1552718
• 负责人: Eric Brustad
• 金额: $ 72.2万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1552718&HistoricalAwards=false
• 关键词: CAREER; Expanding; Protein; Chemistry; through

Expanding Protein Chemistry through the Evolution of Orthogonal Non-natural Heme:Enzyme PairsWith this CAREER award, the Chemistry of Life Processes Program in the National Science Foundation's Division of Chemistry is funding Dr. Eric Brustad from the University of North Carolina at Chapel Hill to expand the possibilities for protein (enzyme) engineers to create new and versatile catalysts for chemical synthesis. Such protein engineering is of growing importance to academic and pharmaceutical research due to the mild and green conditions under which the biological world operates. Therefore the goal of this work is to combine engineering and evolution to create enzyme scaffolds that assemble in cells and which can incorporate building blocks not typically found in nature. This research expands the chemical diversity and reaction landscape available to protein engineers and generates new tools with broad applications in biotechnology and biomedical imaging, as well as chemical synthesis. Dr. Brustad has developed a symposium for biotechnology research to foster communication and collaboration between local scientists and students. This symposium is now expanded to promote diversity by reaching out to Historically Black Colleges and Universities (HBCUs). Dr. Brustad is also developing a hands-on educational modules for K-12 populations in rural North Carolina, serving primarily young African American and Hispanic populations.This research introduces a new paradigm for the construction of non-natural metallocofactor proteins in cells through the creation of orthogonal heme/enzyme pairs. An orthogonal heme:enzyme pair is composed of two key components: 1) a non-natural, synthetic cofactor that is similar to heme but does not interact with existing heme enzymes, and 2) an evolved heme-binding protein that recognizes only the non-natural cofactor. This "orthogonality" creates a new cofactor:enzyme system that can interact self-sufficiently in cells without disrupting endogenous cofactors or enzymes. A significant advantage of this approach is that it simply builds on (and adapts to) existing cofactor-binding motifs in nature to construct binding sites for metals and cofactor catalysts not typically found in nature. Scaffolds identified from this study provide robust and systematically adaptable artificial metalloproteins for green chemical synthesis as well as metal-based imaging applications. In addition, this program is focused on broadening the impact of biotechnology research by expanding education outreach to young underrepresented minorities and undergraduates students as well as fostering increased scientific communication among research institutions across the state of North Carolina.

通过正交非天然血红素的进化扩展蛋白质化学：酶对有了这个职业奖，美国国家科学基金会化学部的生命过程化学计划正在资助北卡罗来纳州大学查佩尔山的Eric Brustad博士，以扩大蛋白质（酶）工程师创造化学合成新的多功能催化剂的可能性。由于生物界的温和和绿色条件，这种蛋白质工程对学术和药物研究越来越重要。因此，这项工作的目标是将联合收割机工程和进化相结合，以创造在细胞中组装的酶支架，并且可以结合自然界中通常不存在的构建块。这项研究扩展了蛋白质工程师可用的化学多样性和反应景观，并产生了在生物技术和生物医学成像以及化学合成中具有广泛应用的新工具。Brustad博士为生物技术研究开发了一个研讨会，以促进当地科学家和学生之间的沟通和合作。这次研讨会现在扩大到通过接触历史上的黑人学院和大学（HBCUs）促进多样性。 Brustad博士还为北卡罗来纳州农村的K-12人群开发了一个实践教育模块，主要服务于年轻的非洲裔美国人和西班牙裔人群。这项研究通过创建正交血红素/酶对，为在细胞中构建非天然金属辅因子蛋白引入了一种新的范式。正交血红素：酶对由两个关键组分组成：1）类似于血红素但不与现有血红素酶相互作用的非天然合成辅因子，以及2）仅识别非天然辅因子的进化的血红素结合蛋白。这种“正交性”创造了一种新的辅因子：酶系统，可以在细胞中自给自足地相互作用，而不破坏内源性辅因子或酶。这种方法的一个显著优点是，它简单地建立在（和适应）现有的辅因子结合基序在自然界中构建的金属和辅因子催化剂的结合位点通常不会在自然界中发现。从这项研究中确定的支架提供了强大的和系统的适应性人工金属蛋白的绿色化学合成以及基于金属的成像应用。此外，该计划的重点是扩大生物技术研究的影响，通过扩大教育推广到年轻的代表性不足的少数民族和本科生，以及促进整个北卡罗来纳州的研究机构之间的科学交流。