Understanding and engineering hydrophobin self-assembly

蛋白质自组装在自然界中发挥着重要作用，为生物材料的开发提供了新的机遇。一类自组装蛋白是疏水蛋白，它是丝状真菌为改变环境而产生的小球状蛋白。分泌的疏水蛋白就像清洁剂一样，可以改变形状形成小棒，这是一种非常耐用和防水的涂层，覆盖在空气中的菌丝和孢子上。疏水性化合物的不同寻常的生化特性引起了人们对开发它们作为乳化剂、泡沫稳定剂和保护和修饰表面的剂的商业应用的兴趣。***目前，对疏水分子的性质及其如何形成小棒缺乏全面的、基于分子的理解。只有少数高分辨率的疏水蛋白结构存在，但这些疏水蛋白在序列和结构上如此多样化，以至于很难将这些疏水蛋白的序列和结构特性与它们的功能联系起来。在与艾玛·马斯特博士（UofT）的合作下，我已经开始描述一个新的疏水分子亚类的结构和功能。通过研究这些疏水蛋白，我将把疏水蛋白的序列特征与它们的功能特性联系起来。具体而言，本提案的目标是：******1。表征各种疏水分子的结构和功能，并确定影响其性质的因素。* * * 2。确定小棒是如何形成的，并建立小棒结构的工作模型。* * * 3。设计可组装成具有所需性能的小棒的疏水桶。******为了实现这些目标，我将使用结合分子生物学、结构生物学和生物物理学的实验方法。这项工作将有助于我们对蛋白质自组装的了解，并将提供对疏水蛋白性质和功能的机制理解，这对设计它们用于商业应用至关重要。我的研究项目将使学员在分子生物学、核磁共振波谱学、x射线晶体学、圆二色光谱偏振学、原子力显微镜和电子显微镜方面发展技能。这些与目前正在商业化开发的蛋白质相关的不同技能和经验将确保高素质人才在未来生物技术、制药和学术领域的职业生涯中取得成功。

Protein self-assembly plays many essential roles in nature and presents new opportunities to develop biomaterials. One family of self-assembling proteins are hydrophobins, which are small globular proteins produced by filamentous fungi to modify their environment. Secreted hydrophobins behave like detergents and can change shape to form rodlets, which are an extremely durable and water repellent coating over aerial hyphae and spores. The unusual biochemical properties of hydrophobins have generated interest in developing them for commercial applications as emulsifiers, foam stabilizers and as an agent to protect and modify surfaces. ***Currently a comprehensive, molecular-based understanding of the properties of hydrophobins and how they form rodlets is lacking. Only a handful of high-resolution structures for hydrophobins exist, but these are for hydrophobins so diverse in sequence and structure that it is difficult to correlate the sequence and structural properties of these hydrophobins with their function. In collaboration with Dr. Emma Master (UofT) I have begun to characterize the structure and function of a new sub-class of hydrophobins. By studying these hydrophobins I will correlate sequence features of hydrophobins with their functional properties. Specifically, the objectives of this proposal are to: ******1. Characterize the structure and function of diverse hydrophobins and determine what factors influence their properties. ***2. Determine how rodlet formation occurs and build a working model of the rodlet structure. ***3. Design hydrophobins that assemble into rodlets with desired properties.******To address these objectives I will use an experimental approach combining molecular biology, structural biology, and biophysics. This work will contribute to our knowledge of protein self-assembly and will provide a mechanistic understanding of hydrophobin properties and function that is critical to engineer them for commercial applications. My research program will allow trainees to develop skills in molecular biology, nuclear magnetic resonance spectroscopy, X-ray crystallography, circular dichroism spectropolarimetry, atomic force microscopy, and electron microscopy. These diverse skills and and experience working with a protein that is currently being developed for commercialization will ensure the success of highly qualified personnel in their future careers in the biotechnology, pharmaceutical and academic sectors.