CAREER: Modular Protein Origami to Build Genetically Programmable Biomaterials

第1部分：非技术概述大自然在纳米尺度上构建生物材料，为生命执行基本功能。就像折纸艺术一样，蛋白质等生物分子折叠并组装成各种形状的小尺寸材料。数十亿年的生命进化已经发展出这种复杂的方法，在每个生命系统中都能制造出非常精确和微小的材料。然而，理解自然的方式并利用它来构建可以编程各种生物功能的人工材料尚未接近实现。这个项目旨在了解一类短的、模块化的螺旋状蛋白质工具，称为线圈，如何折叠和相互作用，并利用蛋白质工具进行分子折纸，以创造纳米级的生物材料。首席研究员将研究和教育工作结合起来，研究盘绕状蛋白质之间的相互作用及其折叠成折纸形状。蛋白质折纸可以用于开发具有可编程复杂功能的生物材料，这可以使前所未有的技术解决生物学和医学问题，并有助于生物制造和医疗保健的进步。利用微生物的遗传操作来产生蛋白质折纸材料，然后利用生物物理表征和计算工具揭示它们的形状和功能。与大学研讨会合作，创建教育模块和外展活动，以产生对蛋白质材料的兴趣，并吸引各级学生，重点是如何设计和制作蛋白质折纸。研究整合活动提供实验室经验，以协助培养本科生和研究生，包括工程专业的女性和历史上代表性不足的学生。本项目旨在了解复杂系统中的可编程蛋白质组装，称为模块化蛋白质折纸，并为具有可定制尺寸，形状和遗传可编程功能的生物纳米材料的设计建立一个强大的框架。卷曲卷曲的蛋白质基序作为通用的模块化工具包，具有特异性、可控和正交的蛋白质-蛋白质相互作用，用于创建定义良好的具有生物功能的蛋白质折纸纳米结构。首席研究员将研究和教育工作结合起来，以全面了解(i)以复杂的折纸拓扑排列的卷曲卷曲蛋白质的相互作用模块性，（ii）如何控制蛋白质折纸并用于细胞中的编程功能，以及（iii）通过细胞表面的模块化蛋白质折纸微生物组装的遗传可编程性。为了实现这一目标，重组蛋白合成的实验技术以及小角度散射和荧光显微镜等蛋白质表征工具与计算结构建模技术相结合。这一结果填补了利用盘绕式蛋白质基序设计生物功能蛋白质纳米材料的关键知识空白，用于从合成生物学到医学等领域的技术开发。教育模块和外展活动为化学工程专业的学生提供了蛋白质材料设计和工程的基础知识，以及蛋白质合成、建模和表征的技术。这些活动吸引了邻近社区中代表性不足的工科学生和K-12学生，重点是在蛋白质折纸设计和制造方面的实践经验。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

PART 1: NON-TECHNICAL SUMMARYNature builds biological materials at the scale of nanometers that perform essential functions for life. Like the paper-folding artwork known as origami, biological molecules such as proteins fold and assemble into small-scale materials in various shapes. The evolution of life for billions of years has developed this sophisticated method to build remarkably precise and tiny materials in every living system. However, understanding nature’s way and exploiting it to build artificial materials that can program various biological functionalities is not yet close to realization. This project aims to understand how a class of short, modular helical protein tools, called coiled-coils, fold and interact with each other and make molecular origami using the protein tools to create nanometer-scale biomaterials. The principal investigator integrates research and education efforts to study the interactions between the coiled-coil proteins and their folding into origami shapes. Protein origami can be used to develop biomaterials with programmable complex functionalities, which can enable unprecedented technologies to solve problems in biology and medicine and contribute to the advancement in biomanufacturing and healthcare. Genetic manipulations of microorganisms are utilized to generate protein origami materials, followed by revealing their shapes and functions using biophysical characterization and computational tools. Education modules and outreach activities, in partnership with university workshops, are created to generate interest in protein materials and engage students of all levels, with a focus on how to design and make protein origami. Research integration activities provide lab experiences to assist the training of undergraduate and graduate students, including women in engineering and historically underrepresented students.PART 2: TECHNICAL SUMMARYThis project aims to understand the programmable protein assembly in complex systems, termed modular protein origami, and to build a robust framework for the design of biological nanomaterials with customizable sizes, shapes, and genetically programmable functionalities. Coiled-coil protein motifs that serve as versatile and modular toolkits with specific, controllable, and orthogonal protein-protein interactions are used to create well-defined protein origami nanostructures that perform biological functions. The principal investigator integrates research and education efforts to gain a comprehensive understanding of (i) the interaction modularity of coiled-coil proteins arranged in complex origami topologies, (ii) how the protein origami is controlled and used to program functionalities in cells, and (iii) genetic programmability of microbial assembly through modular protein origami on cell surfaces. To accomplish the objectives, experimental techniques for recombinant protein synthesis as well as protein characterization tools such as small-angle scattering and fluorescent microscopy are utilized in combination with computational structure modeling techniques. The outcome fills the critical knowledge gap in designing biofunctional protein nanomaterials using the coiled-coil protein motifs for technology development in fields ranging from synthetic biology to medicine. The education modules and outreach activities offer students in chemical engineering exposure to the basics of protein material design and engineering as well as techniques for protein synthesis, modeling, and characterization. The activities engage underrepresented engineering students and K-12 students in the neighboring community with a focus on hands-on experiences in protein origami design and fabrication.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.