CAREER: Molecular Engineering of Bio-orthogonal Stabilized Alpha Helices

1553860格雷格·瑟伯新颖的分子工程战略正在迅速推动独特的生物启发分子的开发，用于材料、分离、药物和医学成像剂应用。然而，这些天然分子经常在使用它们所需的条件下降解，例如在提纯产品时的酸性或碱性pH溶液中，或者在化合物达到治疗目标之前被酶降解的体内。通过使用合成的有机分子来稳定这些生物材料，可以设计出具有巨大复杂性和高度稳定性的试剂。此外，通过选择适当的化学技术，可以开发出高通量的方法，从数百万化合物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基本性质的基础研究，并使应用研究能够开发出适用于不同应用的新型化合物。这项职业计划将结合新的合成连接物、非天然氨基酸(NNAA)的掺入和生物正交化学与定向进化来设计具有独特靶向特性的稳定螺旋。这项建议的目标1将表征一系列新的叠氮-炔稳定的α螺旋，这些螺旋具有独特的化学、成像和药代动力学性质，并与细胞表面定向进化兼容。这种安排只需要在肽骨架中的一种类型的NNAA，以降低复杂性并提高整体掺入效率。较长的连接物可以设计成带有功能性手柄，用于附着到珠子、荧光染料、放射性标记或药代动力学修饰剂上。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。该提案的目标2将利用生物正交化学来选择新的螺旋。引入了一种位点特异性连接，以选择在连接到接头时保持结合的分子。这项建议将通过筛选数百万种用于新疗法、显像剂和分离组件的新型稳定螺旋来缓解目前的开发瓶颈。工程功能多样化的稳定多肽和蛋白质结构将影响包括化学工业、生物医学工程领域、制药工业和医学界在内的多个领域。为了增加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外展计划。与来自以代表性不足为主的背景的学生的多重互动促进了与分子工程研究直接相关的动手活动。这些学生将访问密歇根大学进行互动课程，以增进他们对科学、技术、工程和数学(STEM)领域的理解，并鼓励他们追求科学、技术、工程和数学(STEM)领域。这一职业奖项由CBET部门的生物技术和生化工程项目获得，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.