Protein Folding, Function and Engineering

蛋白质是精致的分子机器--它们执行广泛的功能，从充当催化剂，到专门识别和结合各种分子，再到结构支持角色。线性蛋白质链中氨基酸的顺序，即初级序列，编码蛋白质如何折叠成其功能性的“天然”结构。尽管进行了广泛的研究，但初级序列决定蛋白质折叠和功能的机制细节仍然不清楚。确定这些机制非常重要，因为这些知识最终将使我们能够准确预测任何蛋白质在自然过程、疾病和生物技术中的行为。它将使我们有能力为任何想要的应用设计或设计蛋白质！ 天然蛋白质和工程蛋白质被广泛用于研究和工业，并正在通过它们在材料科学、传感器和药物等方面的应用来改变生物技术。蛋白质序列、结构和功能知识的爆炸性增长以及蛋白质的计算建模为准确阐明序列如何决定折叠和功能奠定了基础。例如，我们开发并展示了一种成功的序列生物信息学和建模方法来设计具有所需结构(三重对称超折叠)和功能(三价碳水化合物结合)的蛋白质(称为ThreeFoil)。我们展示了ThreeFoil在恶劣条件下的超级稳定性，并定义了一种通用机制，解释了大量蛋白质的这种罕见和理想的特性。我们对ThreeFoil和其他蛋白质的发现(包括我们为系统地提高我们的知识而设计的大量突变体)是通过一系列实验(光谱学、热量计、显微镜)和计算方法/工具获得的。细菌通常用于大规模生产蛋白质，这些蛋白质可能正确折叠和溶解，也可能错误折叠成不可溶(但有时仍具有功能)的聚集结构，称为包涵体(IBS)。我们将扩展我们对IB形成的分子机制的最初令人兴奋的发现，以便更好地预测和控制IB的形成和性质。总而言之，我们的研究将促进对蛋白质如何工作以及如何将其用于各种应用的了解；它将为许多高素质的人员提供有价值的跨学科协作和国际培训，以开发蛋白质，从而进一步造福社会。

Proteins are exquisite molecular machines-they perform a vast range of functions, from acting as catalysts, to specifically recognizing and binding all kinds of molecules, to structural support roles. The order of amino acids in the linear protein chain, i.e. the primary sequence, encodes how the protein folds into its functional “native” structure. Despite extensive study, the details of the mechanisms by which primary sequence determines protein folding and function remain obscure. Defining these mechanisms is of tremendous importance because, ultimately, such knowledge will give us the ability to accurately predict the behaviour of any protein, in natural processes, in disease, and in biotechnology. It will give us the ability to engineer or design proteins for any desired application! Natural and engineered proteins are widely used in research and industry and are transforming biotechnology through their applications in material science, sensors and as drugs, for example. Explosive growth in knowledge of protein sequences, structures and functions and in computational modeling of proteins have set the stage for elucidating exactly how sequence determines folding and function. For example, we developed and demonstrated a successful sequence bioinformatics and modelling method to engineer a protein (called ThreeFoil) with a desired structure (three-fold symmetric superfold) and function (trivalent carbohydrate binding). We showed ThreeFoil is super stable against harsh conditions, and defined a general mechanism that explains this rare and desirable property for a great many proteins. Our discoveries for ThreeFoil and other proteins (including numerous mutants which we design to systematically advance our knowledge) are obtained using a battery of experimental (spectroscopy, calorimetry, microscopy) and computational methods/tools. Bacteria are commonly used for the large scale production of proteins, which may be correctly folded and soluble, or misfolded into insoluble (but still sometimes functional) aggregated structures called inclusion bodies (IBs). We will extend our initial exciting discoveries of the molecular mechanisms of IB formation, in order to better predict and control their formation and properties. Collectively, our research will advance knowledge of how proteins work and how they can be harnessed for myriad applications; it will provide valuable collaborative interdisciplinary and international training for many highly qualified personnel in developing proteins for further great benefit to society.