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阳离子共轭聚合物引发分子信标适体构象变化用于高灵敏和选择性钾离子检测。

Cationic conjugated polyelectrolytes-triggered conformational change of molecular beacon aptamer for highly sensitive and selective potassium ion detection.

机构信息

Department of Nano Fusion Technology, Pusan National University, Miryang 627-706, Republic of Korea.

出版信息

J Am Chem Soc. 2012 Feb 15;134(6):3133-8. doi: 10.1021/ja210360v. Epub 2012 Feb 7.

DOI:10.1021/ja210360v
PMID:22280349
Abstract

We demonstrate highly sensitive and selective potassium ion detection against excess sodium ions in water, by modulating the interaction between the G-quadruplex-forming molecular beacon aptamer (MBA) and cationic conjugated polyelectrolyte (CPE). The K(+)-specific aptamer sequence in MBA is used as the molecular recognition element, and the high binding specificity of MBA for potassium ions offers selectivity against a range of metal ions. The hairpin-type MBA labeled with a fluorophore and quencher at both termini undergoes a conformational change (by complexation with CPEs) to either an open-chain form or a G-quadruplex in the absence or presence of K(+) ions. Conformational changes of MBA as well as fluorescence (of the fluorophore in MBA) quenching or amplification via fluorescence resonance energy transfer from CPEs provide clear signal turn-off and -on in the presence or absence of K(+). The detection limit of the K(+) assays is determined to be ~1.5 nM in the presence of 100 mM Na(+) ions, which is ~3 orders of magnitude lower than those reported previously. The successful detection of 5'-adenosine triphosphate (ATP) with the MBA containing an ATP-specific aptamer sequence is also demonstrated using the same sensor scheme. The scheme reported herein is applicable to the detection of other kinds of G-rich aptamer-binding chemicals and biomolecules.

摘要

我们通过调节分子信标适体(MBA)与阳离子共轭聚合物(CPE)之间的相互作用,在水中对过量钠离子表现出高灵敏度和选择性的钾离子检测。MBA 中的钾离子特异性适体序列用作分子识别元件,MBA 对钾离子的高结合特异性提供了对一系列金属离子的选择性。发夹型 MBA 在两端标记荧光团和猝灭剂,在不存在或存在 K(+)离子时,通过与 CPEs 形成复合物,经历构象变化(开链形式或 G-四链体)。MBA 的构象变化以及荧光(MBA 中的荧光团)猝灭或通过 CPEs 从荧光共振能量转移的放大,在存在或不存在 K(+)时提供了明显的信号关闭和开启。在存在 100 mM Na(+)离子的情况下,K(+)测定的检测限被确定为约 1.5 nM,比以前报道的低约 3 个数量级。使用相同的传感器方案还成功地检测了含有 ATP 特异性适体序列的 MBA 对 5'-三磷酸腺苷(ATP)的检测。本文报道的方案适用于检测其他种类的富含 G 的适体结合化学物质和生物分子。

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