A water-soluble conjugated polymer for protein identification and denaturation detection.
A water-soluble conjugated polymer for protein identification and denaturation detection.
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DOI：
10.1002/asia.201000363
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发表时间：
2010-12
期刊：
影响因子：
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通讯作者：
Qingling Xu;Chunxian Wu;Chunlei Zhu;Xinrui Duan;Libing Liu;Yuchun Han;Yilin Wang;Shu Wang
中科院分区：
文献类型：
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作者：
Qingling Xu;Chunxian Wu;Chunlei Zhu;Xinrui Duan;Libing Liu;Yuchun Han;Yilin Wang;Shu Wang
在蛋白质组学、医学诊断学和生物学领域中，快速且灵敏地检测蛋白质和蛋白质变性的方法变得越来越需要。在本文中，我们已经报道了一种新的阳离子水溶性共轭聚合物的合成，含有芴和二烯部分的骨干（PFDE）的蛋白质识别传感阵列的PFDE解决方案，在不同的离子强度使用线性判别分析技术（LDA）。PFDE可以通过静电和/或疏水相互作用与蛋白质形成复合物，并表现出不同的荧光响应。三个主要因素有助于PFDE的荧光响应，即，蛋白质表面上的净电荷密度，蛋白质的疏水性，和金属蛋白质的特性。蛋白质的变性也可以使用PFDE作为荧光探针来检测。用动态光散射（DLS）和等温滴定量热法（ITC）研究了PFDE与蛋白质的相互作用。与其他基于共轭聚合物的方法相比，我们的方法避免了一系列猝灭剂或染料标记的受体或蛋白质底物的合成，这显著降低了成本和合成复杂性。我们的方法提供了一个简单，快速，无标记的方式基于非特异性相互作用诱导的扰动PFDE荧光响应的蛋白质鉴定和变性检测的应用前景。
Rapid and sensitive methods to detect proteins and protein denaturation have become increasingly needful in the field of proteomics, medical diagnostics, and biology. In this paper, we have reported the synthesis of a new cationic water-soluble conjugated polymer that contains fluorene and diene moieties in the backbone (PFDE) for protein identification by sensing an array of PFDE solutions in different ionic strengths using the linear discriminant analysis technique (LDA). The PFDE can form complexes with proteins by electrostatic and/or hydrophobic interactions and exhibits different fluorescence response. Three main factors contribute to the fluorescence response of PFDE, namely, the net charge density on the protein surface, the hydrophobic nature of the protein, and the metalloprotein characteristics. The denaturation of proteins can also be detected using PFDE as a fluorescent probe. The interactions between PFDE and proteins were also studied by dynamic light scattering (DLS) and isothermal titration microcalorimetry (ITC) techniques. In contrast to other methods based on conjugated polymers, the synthesis of a series of quencher or dye-labeled acceptors or protein substrates has been avoided in our method, which significantly reduces the cost and the synthetic complexity. Our method provides promising applications on protein identification and denaturation detection in a simple, fast, and label-free manner based on non-specific interaction-induced perturbation of PFDE fluorescence response.