CAREER: Functional Fouling of Surfaces by Interfacial Silk Fibroin Self-Assembly

非技术总结水中蛋白质随机附着在附近表面的倾向是自然界中普遍存在的现象。这种现象被称为“污垢”，通常被认为是一个令人困惑的问题，可能会导致一系列问题，如生物医学植入物的临床失败和海洋生物在船体上的不良附着。然而，该项目旨在利用蛋白质污垢的持久性作为创建多功能表面的新的、高度通用的方法的基础。利用丝素蛋白，一种源自天然丝素主要成分或受其启发的生物相容性蛋白质，该项目将开发出在各种表面上创建坚固的纳米薄涂层的方法，而不需要危险的化学品或复杂的仪器。为了实现这一目标，该项目将寻求从根本上了解丝素蛋白在自组装过程中的污染行为--这是一种聚集过程，是昆虫和蜘蛛纺丝的基础。通过结合合成生物学和表面敏感材料表征技术，该项目还将揭示自然界中的蛋白质和其他生物大分子如何能够在没有特定化学反应的情况下顽强地附着在表面上的基本见解。此外，该项目将开发一种生物启发战略，通过该战略，表面的功能，如生物兼容性或治疗活性，可以被修改，以应用于人类医疗保健和可持续发展。为了方便向全球公众传播这一项目的复杂科学发现，梅里安视觉交流工作室将在伦斯勒理工学院成立。这个配备了技术的工作空间将使学生能够制作引人注目的可视化教育工具，如2D/3D图形和动画。作为这项视觉交流工作的一部分，该项目还将支持一个由艺术和工程专业的学生组成的跨学科团队来创建一款互动视频游戏，直观地教授自然蛋白质组装和污垢现象的基本概念。技术总结该提案旨在开发一种强大的自下而上的策略，以创建多功能薄膜涂层，利用丝素蛋白在固-液界面的独特自组装行为。这种吸附与蛋白质-蛋白质组装同时发生的界面行为，是一种健壮的“污垢”，尽管是自然生物大分子粘合的一个重要方面，但迄今为止还没有得到充分的研究。此外，虽然蛋白质对表面的污染一直被认为是一个不受欢迎但不可避免的问题，但界面丝素自组装可以作为一种通用的非共价方法，在温和的生物相容条件下修饰各种表面，而不需要复杂的化学反应或先进的仪器。这个项目将通过1)将丝素蛋白在高和低表面能底物上的吸附与可调的溶液相组装现象联系起来，2)确定蛋白质序列和尺寸对使用从头设计的重组丝素蛋白形成涂层的作用，3)可视化纳米级底物形貌对涂层形成的影响，以及4)探索酶和生长因子与丝素蛋白的共同组装，作为一种创建具有治疗活性的表面的方法，来寻求通过界面蛋白自组装获得附着的和无缺陷的涂层的基本原理。该项目将从蛋白质-蛋白质和蛋白质-表面相互作用的角度为网络形成蛋白(如丝素蛋白)的界面组装生成一个全面的模型。此外，这项工作将建立一种新的非共价方法，使表面功能化，用于各种材料应用。为了促进将拟议研究的复杂科学发现传播给广大的非专业受众，该项目将在伦斯勒理工学院建立“梅里安视觉交流工作室”，这将是一个配备技术的工作空间，使学生能够制作引人注目的视觉教育工具，如2D/3D图形和动画。该项目还将支持一个由艺术和工程本科生组成的跨学科团队，以创建一个互动视频游戏，教授与拟议研究相关的蛋白质吸附和组装的基本概念，用于全球外展。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

NON-TECHNICAL SUMMARYThe tendency of proteins in water to adhere randomly to nearby surfaces is a ubiquitous phenomenon in nature. This phenomenon, known as “fouling,” is usually considered a confounding problem which can lead to a wide range of issues, such as clinical failure of biomedical implants and undesirable attachment of marine organisms to ship hulls. However, this project aims to leverage the persistence of protein fouling as the basis of a new, highly versatile approach for creating multifunctional surfaces. Using silk fibroins, a class of biocompatible proteins derived from or inspired by the main component of natural silk, this project will develop methods for creating robust nanometer-thin coatings on a variety of surfaces without requiring hazardous chemicals or complex instrumentation. Towards this goal, the project will seek to gain a fundamental understanding of the fouling behavior of silk fibroins as they undergo self-assembly – an aggregation process which underlies the spinning of silk threads by insects and spiders. Through a combination of synthetic biology and surface-sensitive material characterization techniques, this project will also reveal fundamental insights about how proteins and other biomacromolecules in nature are able to adhere tenaciously to surfaces without specific chemical reactions. Furthermore, this project will develop a bio-inspired strategy by which the function of surfaces, such as biocompatibility or therapeutic activity, can be modified for applications in human healthcare and sustainability. To facilitate the dissemination of complex scientific findings from this project to a global public audience, the Merian Studio for Visual Communication will be founded at Rensselaer Polytechnic Institute. This technology-equipped workspace will enable students to produce compelling visual educational tools, such as 2D/3D graphics and animations. As part of this visual communication effort, the project will also support an interdisciplinary team of art and engineering students to create an interactive video game that intuitively teaches the fundamental concepts of natural protein assembly and fouling phenomena.TECHNICAL SUMMARYThis proposal aims to develop a powerful bottom-up strategy for creating multifunctional thin-film coatings that leverages the unique self-assembly behavior of silk fibroins at solid-liquid interfaces. Such interfacial behavior, where adsorption occurs concurrently with protein-protein assembly, is a type of robust “fouling” which has not been sufficiently studied to date, despite being an important aspect underlying natural biomacromolecular adhesion. Furthermore, whereas fouling of surfaces by proteins has long been considered an undesirable but unavoidable problem, interfacial silk fibroin self-assembly can be used as a versatile non-covalent approach for modifying a variety of surfaces under mild, biocompatible conditions without complex chemistries or advanced instrumentation. This project will seek to gain a fundamental understanding of the growth of adherent and defect-free coatings via interfacial protein self-assembly by 1) relating silk fibroin adsorption on high and low surface energy substrates to tunable solution-phase assembly phenomena, 2) determining the role of protein sequence and size on coating formation using de novo designed recombinant silk fibroins, 3) visualizing the effects of nanoscale substrate topography on coating formation, and 4) exploring co-assembly of enzymes and growth factors with silk fibroins as a one-pot method for creating therapeutically active surfaces. The project will generate a comprehensive model for interfacial assembly of network-forming proteins, such as silk fibroins, in terms of protein-protein and protein-surface interactions. Moreover, this work will establish a new non-covalent approach towards functionalizing surfaces for a variety of materials applications. To facilitate the dissemination of complex scientific findings from the proposed research to a broad lay audience, the project will establish the “Merian Studio for Visual Communication” at Rensselaer Polytechnic Institute, which will be a technology-equipped workspace enabling students to produce compelling visual educational tools, such as 2D/3D graphics and animations. The project will also support an interdisciplinary team of art and engineering undergraduate students to create an interactive video game that teaches fundamental concepts in protein adsorption and assembly related to the proposed research for use in global outreach.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.