Department of Chemistry, Beijing Normal University, Beijing, 100875, China.
Analyst. 2013 Feb 21;138(3):856-62. doi: 10.1039/c2an36564b. Epub 2012 Dec 5.
Potassium ions play diverse roles in biological processes, and abnormal K(+) levels are the hallmarks of diseases. However, the potential clinical application of the developed DNA-based K(+) sensors remains a challenge due to the presence of Pb(2+) in blood samples. In this contribution, a novel colorimetric potassium sensing assay that functions in the presence of Pb(2+) is reported. This approach is based on conformational switching of a hairpin DNA structure to a G-quadruplex. Specifically, the hairpin DNA containing G-rich aptamer T30695 is exposed to successive amounts of Na(+), Pb(2+) and K(+). These cations induce formation of the corresponding metal-stabilized G-quadruplex, which acts as DNAzyme (with hemin as a cofactor) for the catalytic oxidation of ABTS by H(2)O(2). Importantly, studies presented here show that K(+) replaces Pb(2+) from the G-quadruplex to form K(+)-stabilized G-quadruplex, which differ in the catalytic behavior. With Pb(2+)-stabilized G-quadruplex as a probe, a highly sensitive and selective colorimetric detection of K(+) is achieved in the presence of Pb(2+) and excessive Na(+) (140 mM) with the detection limit of 1.9 nM. This system represents the first known DNAzyme-based colorimetric K(+) sensor, which works in the presence of Pb(2+). Finally, the sensor is successfully applied for K(+) detection in a real human serum sample containing Pb(2+).
钾离子在生物过程中发挥着多种作用,而异常的 K(+)水平是疾病的特征。然而,由于血液样本中存在 Pb(2+),开发的基于 DNA 的 K(+)传感器的潜在临床应用仍然是一个挑战。在本研究中,报道了一种在存在 Pb(2+)的情况下具有新型比色钾传感测定法的方法。该方法基于发夹 DNA 结构向 G-四链体的构象转换。具体而言,含有富含 G 的适体 T30695 的发夹 DNA 暴露于连续量的 Na(+)、Pb(2+)和 K(+)。这些阳离子诱导相应的金属稳定的 G-四链体的形成,其充当 DNA 酶(以血红素作为辅因子),用于 H(2)O(2)对 ABTS 的催化氧化。重要的是,本文的研究表明,K(+)从 G-四链体中取代 Pb(2+)以形成 K(+)-稳定的 G-四链体,其在催化行为上有所不同。使用 Pb(2+)-稳定的 G-四链体作为探针,在存在 Pb(2+)和过量的 Na(+)(140 mM)的情况下,实现了对 K(+)的高灵敏度和选择性比色检测,检测限为 1.9 nM。该系统代表了第一个已知的基于 DNA 酶的比色 K(+)传感器,其在存在 Pb(2+)的情况下工作。最后,该传感器成功应用于含有 Pb(2+)的真实人血清样本中的 K(+)检测。