Small-molecule sensing and capture by de novo protein receptors followed by fluorescence spectroscopy and mass spectrometry.

蛋白质在自然界中已经有了一系列令人难以置信的多功能结构和功能，我们对它们的理解使新结构的设计成为可能。从头蛋白质设计使蛋白质的创造能够用于工业生物技术或工程，而不受自然界中发现的选择压力的影响。一个蛋白质折叠，卷曲螺旋，是一个成功的从头设计目标的例子。它们的结构可以参数化描述，这使得计算设计和发现新的低聚态在自然界中还看不见。高阶卷曲螺旋具有贯穿其结构的连续孔，称为α-螺旋桶（aHB）。这些aHB已被证明是令人难以置信的热稳定性和耐受的几个突变的管腔定向残基。aHB已经被用于进行简单的化学修饰、完整的催化循环和捕获小分子。后者才刚刚开始探索。在目前的理解中，脂质是aHB最有吸引力的靶点。脂质组可用于通过使用特征性生物标志物来鉴定许多疾病，包括癌症、肥胖症、阿尔茨海默病和糖尿病。处理过的脂质样品通常通过使用质谱结合色谱分离方法进行分析。然而，这种代谢组学研究可能是密集的，需要复杂的仪器与大量的知识来分析结果。在这里，我们的目标是确定特定的aHB：在复杂的流体中的配体结合伙伴，以产生简单的阅读，定量比色诊断测试，通过使用aHB阵列与环境敏感染料串联。aHB本身吸引疏水性化合物，对化合物大小具有一定特异性。增强我们对一般结合如何发生的理解将进一步使肽设计具有独特的结合配偶体类型。特定大分子的差异感测可以实现不同条件的指纹识别以及独特化合物的存在或缺乏。其结果将是一个超稳定，可变和易于阅读的诊断阵列，在工业和医学中的应用。

Proteins already have an incredibly versatile array of structures and functions in nature and our understanding of them has enabled the design of new structures. De novo protein design has enabled the creation of proteins for use in industrial biotechnologies or engineering, free of the selective pressures found in nature. One protein fold, coiled coils, are one such example of successful de novo design targets. Their structure can be described parametrically which has allowed for computational design, and the discovery of new oligomeric states as of yet unseen in nature. Higher order coiled coils possess a continuous pore running through their structure, named a-helical barrels (aHBs). These aHBs have been shown to be incredibly thermostable and tolerable of several mutations in the lumen orientated residues. aHBs have already been exploited to perform simple chemical modifications, full catalytic cycles and the capturing of small-molecules. The latter of which is only just beginning to be explored. At the current understanding, lipids are the most attractive targets for aHBs. The lipidome can be used to identify many diseases through the use of characteristic biomarkers including cancers, obesity, Alzheimer's disease and diabetes. Treated lipids samples are often analysed through the use of mass spectroscopy coupled with a chromatographic separation method. However, such metabolomic studies can be intensive, requiring complex instrumentation with a large perquisite of knowledge to analyse the results. Here, we aim to identify specific aHB:ligand binding partners in complex fluids in order to produce simple to read, quantitative colorimetric diagnostic tests through the use of aHBs arrays in tandem with environment-sensitive dyes. aHBs by their nature attract hydrophobic compounds with some specificity on compound size. Enhancing our understanding of how the general binding occurs would further enable peptide design with unique binding-partner types in mind. The differential sensing of particular macromolecules can enable the fingerprinting of different conditions and the presence, or lack thereof, of unique compounds. The result would be a hyper-stable, mutable and easily readable diagnostic array with applications in industry and medicine.