Understanding and engineering hydrophobin self-assembly

了解和设计疏水蛋白自组装

• 批准号: RGPIN-2017-05338
• 负责人: Langelaan, David
• 金额: $ 1.89万
• 依托单位: Dalhousie University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=656579
• 关键词: Understanding; engineering; hydrophobin; self; assembly

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.

蛋白质自组装在自然界中发挥着重要作用，为生物材料的发展提供了新的机遇。自组装蛋白的一个家族是疏水蛋白，其是由丝状真菌产生以改变其环境的小球状蛋白。分泌的疏水蛋白表现得像洗涤剂，可以改变形状形成棒状物，棒状物是气生菌丝和孢子上极其耐用的防水涂层。疏水蛋白的不寻常的生物化学性质产生了开发它们作为乳化剂、泡沫稳定剂和作为保护和改性表面的试剂的商业应用的兴趣。* 目前，人们对疏水蛋白的性质以及它们如何形成小棒缺乏全面的、基于分子的了解。只有少数的疏水蛋白的高分辨率结构存在，但这些是疏水蛋白的序列和结构如此不同，很难将这些疏水蛋白的序列和结构特性与它们的功能相关联。在与Emma Master博士（UofT）的合作中，我已经开始表征疏水蛋白的一个新亚类的结构和功能。通过研究这些疏水蛋白，我将疏水蛋白的序列特征与它们的功能特性相关联。具体而言，本提案的目标是：*1。表征不同疏水蛋白的结构和功能，并确定影响其特性的因素。***2。确定小棒的形成是如何发生的，并建立小棒结构的工作模型。*3。设计疏水蛋白，组装成具有所需特性的小棒。**为了达到这些目标，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的蛋白质自组装的知识，并将提供疏水蛋白的性质和功能，这是至关重要的工程商业应用的机械理解。我的研究计划将允许学员发展分子生物学，核磁共振光谱学，X射线晶体学，圆二色谱光谱偏振，原子力显微镜和电子显微镜的技能。这些不同的技能和经验与目前正在开发的商业化蛋白质的工作将确保高素质的人员在他们未来的职业生涯中在生物技术，制药和学术部门的成功。