CAREER: Molecular Engineering of Bio-orthogonal Stabilized Alpha Helices

1553860 Thurber， Greg新颖的分子工程策略正在迅速开发独特的生物启发分子，用于材料，分离，药物和医学成像剂应用。然而，这些天然分子通常在其使用所需的条件下降解，例如在净化产品时在酸性或碱性pH溶液中，或在体内酶降解化合物才能达到治疗目标。通过使用合成有机分子来稳定这些生物材料，可以设计出具有高度复杂性和高稳定性的制剂。此外，通过选择合适的化学技术，可以开发出高通量方法，从数以百万计的化合物文库中快速选择有用的分子。这项工作将支持对这些半合成材料的基本特性的基础研究，并使应用研究能够开发用于各种应用的新化合物。该CAREER提案将结合新型合成连接剂、非天然氨基酸（NNAA）掺入、生物正交化学和定向进化来设计具有独特靶向特性的稳定螺旋。本提案的目标1将描述一系列具有独特化学、成像和药代动力学性质的叠氮-炔稳定α螺旋，并与细胞表面定向进化兼容。这种安排只需要肽主链中的一种NNAA，以减少复杂性并提高整体结合效率。较长的连接可以设计与功能手柄连接珠，荧光染料，放射性标签，或药代动力学修饰剂。这种模块化设计使这些分子能够用于多种应用，包括分离，细胞内靶向和成像。本课题的目的二是利用生物正交化学选择新型螺旋。一个位点特异性的链接被引入来选择分子，当连接到连接器上时将保持结合。该提案将通过筛选数百万种新型稳定螺旋来缓解当前的发展瓶颈，用于新疗法、显像剂和分离成分。工程功能多样化的稳定肽和蛋白质结构将影响化学工业、生物医学工程、制药工业和医学界等多个领域。为了培养高素质和多样化的工程专业学生来完成这些工作，将为底特律地区资源不足的当地学校的学生创建一个以科学为重点的拓展项目。与主要来自代表性不足背景的学生进行多次互动，促进与分子工程研究直接相关的实践活动。这些学生将访问密歇根大学，参加互动会议，以增强他们对科学的理解，并鼓励他们从事科学、技术、工程和数学（STEM）领域的研究。该职业奖由CBET部门的生物技术和生化工程项目颁发，由分子和细胞生物科学部的系统和合成生物学项目共同资助。

1553860 Thurber, Greg Novel molecular engineering strategies are rapidly enabling the development of unique bio-inspired molecules for materials, separations, drug, and medical imaging agent applications. However, these natural molecules often degrade at the conditions necessary for their use, such as in acidic or basic pH solutions when purifying a product or in the body where enzymes degrade the compounds before they can reach their therapeutic target. By using synthetic organic molecules to stabilize these biological materials, the agents can be engineered with both vast complexity and high stability. Furthermore, by choosing an appropriate chemical technique, high-throughput methods can be developed to rapidly select useful molecules from libraries of millions of compounds. This work will support basic research into the fundamental properties of these semi-synthetic materials and enable applied research to develop novel compounds for diverse applications. This CAREER proposal will combine novel synthetic linkers, non-natural amino acid (NNAA) incorporation, and bio-orthogonal chemistry with directed evolution to engineer stabilized helices with unique targeting properties. Aim 1 of this proposal will characterize a novel series of azide-alkyne stabilized alpha helices that possess unique chemical, imaging, and pharmacokinetic properties and are compatible with cell surface directed evolution. This arrangement only requires one type of NNAA in the peptide backbone to reduce complexity and increase overall incorporation efficiency. The longer linker can be engineered with a functional handle for attachment to beads, fluorescent dyes, radiolabels, or pharmacokinetic modifiers. This modular design enables these molecules to be used in multiple applications including separations, intracellular targeting, and imaging. Aim 2 of the proposal will capitalize on the bio-orthogonal chemistry to select novel helices. A site-specific linkage is introduced to select for molecules that will retain binding while attached to the linker. This proposal will alleviate the current development bottleneck by screening millions of novel stabilized helices for new therapeutics, imaging agents, and separations components. Engineering functionally diverse stabilized peptide and protein structures will impact several fields including the chemical industry, biomedical engineering field, pharmaceutical industry, and medical community. To grow the pipeline of highly qualified and diverse engineering students to fulfill these jobs, a science-focused outreach program will be created for students from local under-resourced schools in the Detroit area. Multiple interactions with students from predominantly under-represented backgrounds facilitate hands-on activities directly related to molecular engineering research. These students will visit the University of Michigan for interactive sessions to enhance their scientific understanding and encourage them to pursue science, technology, engineering, and mathematics (STEM) fields.This CAREER award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biosciences.