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Ferrocene-peptide adducts for DNA binding: Towards sequence-selective electrochemical DNA sensors

用于 DNA 结合的二茂铁-肽加合物：走向序列选择性电化学 DNA 传感器

基本信息

批准号：
EP/J014672/1
负责人：
Anna Peacock
金额：
$ 12.71万
依托单位：
University of Birmingham
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2012
资助国家：
英国
起止时间：
2012 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FJ014672%2F1
关键词：
Ferrocene peptide adducts DNA binding

项目摘要

DNA is crucial to life on earth, as it contains the genetic information which defines the characteristics of any living thing, from plankton to humans. It is made up of sequences of smaller building blocks, called nucleobases. This sequence defines both the genetic code the DNA contains and also its physical structure, which is most commonly a double helix - a bit like a ladder, coiled into a spiral. Scientists have successfully decoded the entire sequence of human DNA and in turn have been able to link specific genes - discrete stretches of a DNA sequence - to particular diseases. This offers the potential to selectively identify genes which cause disease and offer new therapies. However, as the human genome alone contains around 20,000 genes, it is very important to be able to selectively and reliably recognise and signal the presence of individual genes. The type of molecule or device that allows us to do this is called a Sensor.Sensors typically comprise two parts; recognition units which are highly selective for the desired target, and a reporter group which allows a measureable and quantifiable output. Chemists have had significant success in developing sensors to analyse small molecules, for instance pollutants and drug molecules. However, these molecules typically interact with sensors on the atomic level, whereas DNA and other biomolecules interact at a level 10 times larger than this, so a new class of sensor needs to be developed.Nature uses DNA-binding proteins to "read" the genetic information stored in the DNA sequence. Proteins are long chains of smaller building blocks called amino acids that fold into well-defined structures, which are important for biomolecular recognition. Though these proteins are large and complicated molecules, the majority bind to the major groove of double helix DNA using a relatively small helical sequence, which is cylindrical in shape. It is our intention to capture this DNA binding strength and selectivity in a significantly simpler (easily synthesised) system, by preparing a minimalist protein sequence derived from a DNA-binding helix. This will then be coupled to a chemical reporter group to provide the user with a measurable output sensitive to DNA binding. Due to its attractive properties and ease of modification, ferrocene is our reporter group of choice. We propose to use a core ferrocene unit and couple it to two DNA-binding helices which will be taken from the protein GCN4. This coupling will be achieved through the side chain of cysteine, a natural amino acid which we will introduce into the GCN4 sequence. This synthesis will lead to the development of novel, sequence-selective DNA biosensors, and provide an approach which is more widely applicable.These miniature ferrocene-protein biosensors would offer advantages over current DNA sensors as they would be capable of selecting for both particular sequences and for double helix DNA. Other advantages would include being able to mimic the biological function of the DNA binding protein from which the sensor is derived, while providing a signal output, allowing us to monitor important biological processes taking place in real time.These new sensors will drive increased understanding of how nature recognises genes within DNA, and in doing so will contribute greatly to efforts to develop new medical treatments and diagnostics.

DNA对地球上的生命至关重要，因为它包含的遗传信息决定了从浮游生物到人类的任何生物的特征。它是由称为核碱基的更小的构建块序列组成的。这个序列既定义了DNA包含的遗传密码，也定义了它的物理结构，最常见的是双螺旋结构——有点像梯子，盘绕成螺旋状。科学家们已经成功地解码了人类DNA的整个序列，进而能够将特定的基因——DNA序列的离散片段——与特定的疾病联系起来。这就有可能选择性地识别导致疾病的基因，并提供新的治疗方法。然而，由于人类基因组包含大约20,000个基因，因此能够有选择地、可靠地识别和标记单个基因的存在是非常重要的。允许我们这样做的分子或设备的类型被称为传感器。传感器通常由两部分组成；识别单元是高度选择性的期望目标，和一个报告组，允许一个可测量和可量化的输出。化学家们在开发用于分析小分子的传感器方面取得了重大成功，例如污染物和药物分子。然而，这些分子通常在原子水平上与传感器相互作用，而DNA和其他生物分子在比这大10倍的水平上相互作用，因此需要开发一种新型传感器。大自然利用DNA结合蛋白“读取”存储在DNA序列中的遗传信息。蛋白质是由被称为氨基酸的小构件组成的长链，折叠成明确的结构，这对生物分子识别很重要。虽然这些蛋白质是大而复杂的分子，但大多数都是通过一个相对较小的螺旋序列结合到双螺旋DNA的主要凹槽上的，这个螺旋序列是圆柱形的。我们的目的是通过制备来自DNA结合螺旋的极简蛋白序列，在一个更简单（易于合成）的系统中捕获这种DNA结合强度和选择性。然后将其与化学报告组耦合，为用户提供对DNA结合敏感的可测量输出。由于二茂铁具有吸引人的性质和易于改性，因此是我们的报告基团的选择。我们建议使用一个核心二茂铁单元，并将其偶联到两个dna结合螺旋上，这些螺旋将从蛋白质GCN4中取出。这种偶联将通过半胱氨酸侧链实现，这是一种天然氨基酸，我们将引入GCN4序列。这种合成将导致新型的、序列选择性的DNA生物传感器的发展，并提供一种更广泛适用的方法。这些微型二茂铁蛋白生物传感器将比现有的DNA传感器具有优势，因为它们能够选择特定序列和双螺旋DNA。其他优点包括能够模拟传感器的DNA结合蛋白的生物功能，同时提供信号输出，使我们能够实时监测重要的生物过程。这些新型传感器将推动人们进一步了解大自然如何识别DNA中的基因，并在此过程中为开发新的医学治疗和诊断方法做出巨大贡献。