Smart Materials: Development of Protein Recognition Polymers as Novel Surfaces for Protein Crystallisation.

我们提出的研究调查了一种使用新型蛋白质识别聚合物（PRPs）作为诱导蛋白质结晶表面的新方法。蛋白质结晶对人类基因组计划的成功至关重要，该计划为疾病的治疗开辟了令人兴奋的机会。然而，潜在药物的目标往往不是基因本身，而是由这些基因编码的数千种蛋白质。为了理解蛋白质是如何完成各种任务的，我们需要知道这些蛋白质的结构。测定蛋白质结构最有效的技术是x射线晶体学，这需要高质量的蛋白质晶体。目前，世界范围内的结构基因组学/蛋白质组学项目已经开始确定10万种蛋白质的结构，尽管投入了大量的资金和努力，但他们取得的成功有限，因为获得晶体是进展的主要瓶颈。迫切需要新的和改进的方法来促进晶体的生长。问题在于要么根本得不到晶体，要么更令人沮丧——得到的晶体质量不够高，无法进行结构测定。获得好晶体的最终手段是控制它们的孕育阶段，即成核阶段，这是决定整个结晶过程的第一步。这不是一项容易的任务，因此，圣杯是找到一种“通用成核剂”——一种能诱导任何蛋白质结晶的底物。一些物质，如矿物质、人的头发、二氧化硅都被用作核剂，但收效甚微。PRP可以选择性地识别特定的蛋白质，因此很有可能成为理想的成核剂，因为与其使用随机材料作为成核剂，还可以在溶液中为每种蛋白质制备PRP，然后使用该PRP使其结晶。蛋白质结晶研究的结果将使PRPs用于一般蛋白质捕获应用（例如蛋白质提取和蛋白质生物传感）的优化成为可能。PRP技术仍处于起步阶段，目前还不存在适当的表征方法。成核研究将提供一条有趣的途径，以反馈有关交联密度和单体选择等参数是否能导致更好的PRP形成的基本信息。目前，用于蛋白质的生物传感器的开发成本很高，因为它们需要使用昂贵的抗体。由于具有仿生学能力（以及它们作为合成抗体的潜力），PRPs可能为开发用于癌症标志物和生物恐怖主义标志物的新型低成本生物传感器提供了一条途径。

Our proposed research investigates a new approach to the use of a novel class of protein recognition polymers (PRPs) as surfaces for inducing protein crystallisation.Protein crystallisation is vital to the Success of the Human Genome project which has opened up exciting opportunities for the treatment of disease. However, it is often not the genes themselves that are the targets of potential drugs, but the thousands of proteins encoded by these genes. In order to understand how proteins perform their various tasks, we need to know the structure of these proteins. The most effective technique for determining protein structure is X-ray crystallography which requires high quality crystals of the proteins. The current structural genomics/proteomics projects worldwide have set out to determine the structures of 100,000 proteins but in spite of investing considerable funds and effort, they have had limited success because obtaining crystals is a major bottleneck to progress. There is an urgent need for new and improved ways to enable the growth of crystals.The problem lies in either getting no crystals at all, or even more frustrating - getting crystals that are not of a high enough quality to allow structure determination. The ultimate means to obtain good crystals is to control their conception stage, i.e. the nucleation stage which is the first step that determines the entire crystallization process. This is not an easy task hence the Holy grail is to find a 'universal nucleant' - a substrate that would induce crystallization of any protein. Several substances such as minerals, human hair, silicas have been used as nucleants, but with limited success. PRPs can selectively recognize particular proteins and therefore are very likely to serve as the ideal nucleants because rather than use random materials as nucleants, it will be possible to prepare PRPs for each protein while it is in solution and then get it to crystallise using that PRP. The results of the protein crystallisation studies will enable optimization of the PRPs for general protein capture applications (e.g. protein extraction and protein biosensing). The PRP technology is still in its infancy and proper characterisation methods do not currently exist. The nucleation studies will allow an interesting route to the feedback of essential information regarding whether parameters such as crosslinking density and choice of monomers can lead to better PRP formation. Biosensors for proteins are currently expensive to develop because they require the use of expensive antibodies. Because of the biomimicry capabilities (and their potential to act as synthetic antibodies), PRPs potentially offer a route to the development of new low-cost biosensors for cancer markers and bioterrorism markers.