Water-soluble gold nanoclusters prepared by protein-ligand interaction as fluorescent probe for real-time assay of pyrophosphatase activity
Water-soluble gold nanoclusters prepared by protein-ligand interaction as fluorescent probe for real-time assay of pyrophosphatase activity
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DOI：
10.1016/j.bios.2016.04.031
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发表时间：
2016-09-15
影响因子：
12.6
通讯作者：
Chen, Wei
中科院分区：
文献类型：
--
作者：
Deng, Hao-Hua;Wang, Fei-Fei;Chen, Wei
本文报道了一种通过蛋白质-配体相互作用合成水溶性巯基保护金纳米粒子的新方法。以3-巯基丙酸（MPA）为模型配体，牛血清白蛋白（BSA）为模型蛋白，制备了具有强烈橙子黄色荧光发射的水溶性金纳米粒子（BSA/MPA-AuNCs），量子产率为16%.结果表明，该方法制备的纳米金与BSA之间存在疏水相互作用。合成策略，然后成功地扩展到生产水溶性的AuNC保护的其他硫醇盐。利用Fe 3+与BSA/MPA-AuNCs的选择性配位、焦磷酸盐（PPi）与Fe 3+的高亲和力以及PPase对PPi的水解作用，建立了一种灵敏、环保的检测无机焦磷酸酶（PPase）活性的传感系统。在0.1 ~ 3U/L范围内，荧光强度与PPase活性呈良好的线性关系。检出限为0.07 U/L。此外，本文开发的荧光测定法可用于测定真实的生物样品中的PPase活性并评价PPase抑制剂，说明了生物分析的巨大潜力。(C)2016爱思唯尔B. V.保留所有权利。
This paper reports a new and facile method for the synthesis of water-soluble thiolate-protected AuNCs via protein-ligand interaction. Using 3-mercaptopropionic acid (MPA) as a model ligand and bovine serum albumin (BSA) as a model protein, water-soluble AuNCs (BSA/MPA-AuNCs) with intense orange yellow fluorescent emission (quantum yield = 16%) are obtained. Results show that AuNCs produced with this method have hydrophobic interactions with BSA. The synthetic strategy is then successfully extended to produce water-soluble AuNCs protected by other thiolates. Moreover, a sensitive and eco-friendly sensing system is established for detection of the activity of inorganic pyrophosphatase (PPase), which relies on the selective coordination of Fe3+ with BSA/MPA-AuNCs, the higher affinity between pyrophosphate (PPi) and Fe3+, and the hydrolysis of PPi by PPase. A good linearity between the fluorescence intensity and PPase activity within the range from 0.1 to 3 U/L. is found, with a detection limit down to 0.07 U/L. Additionally, the fluorescent assay developed here is utilized to assay the PPase activity in real biological samples and as well as to evaluate PPase inhibitor, illustrating the great potential for biological analysis. (C) 2016 Elsevier B.V. All rights reserved.