Solubilising Insoluble Proteins using Self-Assembling Supramolecular Structures

对于研究分离蛋白的研究人员来说，蛋白质的不溶解性是一个常见的问题，因为分离蛋白表面存在疏水性斑块，在自然环境外会发生错误折叠和/或聚集。由此产生的不溶性材料通常很难（如果不是不可能的话）表征，并且通常不会显示出研究人员着手探索的有用和有趣的特性。因此，将不溶性蛋白质转化为可溶性蛋白质的方法受到高度追捧，因为它们可能允许表征新蛋白质，或者充分稳定酶，以便在非生物环境中利用它们的催化功能，例如在化学制造或生物修复应用中。克服与疏水蛋白相关的溶解度挑战的一个合理方法是将单个蛋白质封装在具有疏水空腔的水溶性胶囊中。这种“隐形涂层”将支持蛋白质暴露其表面疏水残基的自然倾向，同时将蛋白质带入溶液中，从而使酶在水环境中进一步研究或应用。近年来，在开发具有疏水腔的超分子结构方面取得了重大进展，这些超分子结构能够容纳包括生物分子在内的一系列客体分子我们建议研究硅蛋白的封装，硅蛋白3是一种工业上相关的酶，具有较差的溶解度，在自组装金属有机材料中。包封后，酶的活性将被评估，并试图表征的结构，这还有待阐明，将进行。被任命为该职位的学生将从事一个令人兴奋的材料化学和生物化学边界的跨学科项目，并将在蛋白质表达和纯化技术，化学合成，核磁共振分析，光谱学和PXRD方面接受全面培训，并接受质谱方面的高水平培训。

Protein insolubility is a common problem for researchers working with isolated proteins that, due to hydrophobic patches on their surface, misfold and/or aggregate outside of their natural environment. The resulting insoluble materials are frequently hard (if not impossible) to characterise, and often do not display the useful and interesting properties researchers set out to explore. Methodologies that transform insoluble proteins into soluble species are therefore highly sought after as they may allow characterisation of novel proteins, or stabilise enzymes sufficiently for their catalytic function to be harnessed in abiological environments, for example in chemical manufacturing or bioremediation applications. One rational approach to overcoming the solubility challenges associated with hydrophobic proteins is encapsulation of individual proteins within water-soluble capsules that have hydrophobic cavities. Such a 'stealth coating' will support the natural tendency of the protein to expose its hydrophobic residues on its surface while at the same time bringing the protein into solution enabling further study or application of the enzyme in aqueous environments. In recent years significant advances have been made in developing supramolecular structures with hydrophobic cavities that are able to accommodate a range of guest molecules including biomolecules.1,2 We propose investigating the encapsulation of silicatein,3 an industrially relevant enzyme with poor solubility, within self-assembling metal-organic materials. Following encapsulation the activity of the enzyme will be assessed and attempted characterisation of the structure, which has yet to be elucidated, will be undertaken. The student appointed to this position will work on an exciting interdisciplinary project at the boundary of materials chemistry and biochemistry and will be fully trained in protein expression and purification techniques, chemical synthesis, NMR analysis, optical spectroscopy and PXRD as well as receiving high level training in mass spectrometry.