Two-dimensional protein arrays: de novo design and applications

1401835 Baneyx, Francois C.二维（2D）蛋白质阵列对于先进材料、设备和系统的生产具有相当大的兴趣。例如，天然存在的古细菌细胞紫色膜块已被用于光学信息存储；从古细菌和细菌中分离出的二维蛋白质层已被用于生产下一代疫苗，并作为制造纳米结构无机材料的模板。然而，这些技术还没有发展到超出概念证明的程度，因为它们不够强大或可扩展，而且这些系统的几何结构、化学结构或组装都无法从纳米尺度精确控制到中尺度。在这项工作中，建议创建全新一代的自组装2D支架，该支架从头开始设计，以实现下一代先进材料，生物传感器和生物设备。该项目无缝融合了计算，分子生物学，生物化学，生物技术和材料科学，并提供了一个独特的机会，培养学生在真正的跨学科研究。一名代表性不足的本科生将与一名研究生配对，以扩大对科学的参与。这项工作的成果和经验教训将向公众展示，并将开发一个为期一周的自组装模块，纳入该部门的分子工程课程。本文建议通过计算设计一组蛋白质，通过融合来自对称兼容低聚物的原聚物和重新设计单元细胞之间的界面接触，使其能够强大地自组装成二维阵列。我们将描述和优化可扩展形成的二维阵列的自组装条件，在纳米尺度上保持短程顺序，但增长超过数百微米。我们将开发将这些晶格堆叠成周期性3D结构的方法，并通过静电相互作用、表面修饰和结构允许位点内固体结合肽的工程来控制技术相关界面上二维阵列的吸附和取向。最后，我们将探索这些设计二维晶格的潜力，作为无机矿化模板的可调模具，以及作为酶和蛋白质高密度显示和膜蛋白结晶的支架。

1401835 Baneyx, Francois C. Two dimensional (2D) protein arrays are of considerable interest for the production of advanced materials, devices and systems. For instance, the naturally occurring purple membrane patches of archaeal cells have been exploited for optical information storage; 2D protein layers isolated from archaea and bacteria have been used to produce next generation vaccines and as templates for the fabrication of nanostructured inorganic materials. Yet, these technologies have not evolved beyond proof of concept because they are not robust or scalable, and because neither the geometry, nor the chemistry or assembly of these systems can be precisely controlled from the nano- to the mesoscale. In this work, it is proposed to create an entirely new generation of self-assembled 2D scaffolds that have been designed from the ground up to enable the next generation of advanced materials, biosensors and biodevices. The project seamlessly blends computation, molecular biology, biochemistry, biotechnology, and materials science, and provides a unique opportunity to train students in truly interdisciplinary research. One underrepresented undergraduate student will be paired with a graduate student to broaden participation in science. Results and lessons from the proposed work will be presented to the public and a week-long module on self-assembly will be developed for inclusion in the department's Molecular Engineering curriculum. It is proposed to computationally design a family of proteins that will be capable of robust self-assembly into 2D arrays by fusing protomers from symmetry-compatible oligomers and redesigning interfacial contacts between unit cells. We will delineate and optimize self-assembly conditions for scalable formation of 2D arrays retaining short-range order at the nanoscale but growing over hundreds of micrometers. We will develop approaches to stack these lattices into periodic 3D structures, and apply strategies to control the adsorption and orientation of 2D arrays at technological-relevant interfaces through electrostatic interactions, surface modification and the engineering of solid binding peptides within permissive sites of the structure. And finally, we will explore the potential of these designer 2D lattices as tunable molds for templating inorganic mineralization and as scaffolds for high-density display of enzymes and proteins and for membrane protein crystallization.