| 功能肽段介导的铜离子荧光传感器的构建及应用 |
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| 基金项目:国家自然科学基金资助项目( 21775114,81471696) |
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| 中文摘要:铜离子是导致多种疾病(如阿尔茨海默病、朊病毒病和肌萎缩性侧索硬化症等)的重要因素。当前已有针对铜离子检测的相关报道，但基于多肽与铜离子相互作用的荧光传感器鲜有关注。该文依据淀粉样蛋白与铜离子特异性结合的特点，利用已知的淀粉样蛋白组成单元，设计出能与铜离子有序配位的多肽分子(DDAEGHARHCR)。同时，借助于“多肽-铜离子-钙黄绿素”三元竞争体系，构建出铜离子的荧光恢复性传感器。铜离子能有效地猝灭钙黄绿素的荧光信号。而当向钙黄绿素-铜离子体系引入功能肽段时，肽段凭借更强的结合能力将铜离子夺取出来，极大地恢复体系的荧光信号，进而完成对铜离子的传感检测。在对体系的反应温度、反应时间、pH值、钙黄绿素浓度等实验参数进行优化后，实现了对环境水样中铜离子的高灵敏检测。该方法对铜离子检出限为127 nmol/L，线性范围为0.12~13 μmol/L，具有简单、快速、高效、稳定性好等特点，满足环境水样的检测需求，在水体中铜离子含量的监测与分析方面展现出良好的应用潜力。 |
| 中文关键词:多肽 铜离子 金属蛋白 荧光检测 |
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| Construction and Application of a Functional Peptide-mediated Fluorescence Biosensor for Copper Ions Detection |
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| Abstract:Problems of heavy-metal pollution have been drawing more and more attraction of all parts of society.To establish a simple,rapid,stable and efficient detection method for heavy metal ions is an urgent need for environmental stability and life-health.The copper ions(Cu(Ⅱ)) may be an important factor leading to several diseases,including Alzheimer's disease,prion disease and amyotrophic lateral sclerosis.Although some significant researches have been reported in development of chemosensors to detect Cu(Ⅱ),the development of peptide-based fluorescent chemosensors capable of detecting multiple ions has received considerably less attention.The metalloprotein database offers a great possibility for the design and screening of sensing elements in metal ions detection.Through the comparative analysis of the interactions between biomolecules(e-g.proteins,peptides and amino acids) and metal ions,the high specificity and stability of the binding of peptide molecules to metal ions have been determined.In combination with such advantages of peptide molecules as stability,flexibility,and easy modification,a series of analytical studies were conducted on their behaviors with metal ions.This paper was devoted to the construction of a fluorescent Cu(Ⅱ) biosensor based on the specific performance between peptides and Cu(Ⅱ) ions.In view of the flexible design and coordination sites of peptides,the interaction between peptides and Cu(Ⅱ) was deeply researched.A peptide(DDAEGHARHCR) specifically coordinated with Cu2+ was engineered by utilizing the known amyloid protein.And their binding ability and specifity were confirmed by the characterization of MALDI-TOF MS.With the formation of the ternary-competition system “peptides-Cu(Ⅱ)-calcein”,a unique fluorescence biosensor was constructed,thereby achieving the highly sensitive detection of Cu(Ⅱ) in water samples.As is known,Cu(Ⅱ) ions could effectively quench the fluorescence signal of calcein.When the functional peptide was introduced into the calcein-Cu(Ⅱ) system,the peptide with higher binding ability could promptly seize the Cu(Ⅱ) ions,greatly recovering the fluorescence signal of this system.Thus,the effective Cu(Ⅱ) sensing detection could be built.After optimizing the reaction temperature,reaction time,pH and calcein concentration,the detection of Cu(Ⅱ) in environmental water samples was also realized.Results demonstrated that the detection limit of this method for Cu(Ⅱ) was as low as 127 nmol/L,with a linear range of 0.12-13 μmol/L.To ensure the reliabity,the Cu(Ⅱ) concentrations was quantified by atomic absorption spectroscopy(AAS).The high consistency between AAS and our results indicated the accuracy and dependability of the work.Compared with other approaches for Cu2+ detection,the established method exhibited superiorities in simplicity,selectivity,efficiency and stability.Furthermore,the protocol involving the peptides design had a great application potential and a broad development prospect in pratical detection,which also offered a novel idea for the constitution of recognition elements. |
| Key Words:peptides copper ions metalloprotein fluorescence detection |
| 引用本文：李昕翼，冯富岩，周晓东,胡继明.功能肽段介导的铜离子荧光传感器的构建及应用[J].分析测试学报,2021,40(6):907-913. |
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