铽-G-四链体-血红素复合物的过氧化物氧化酶活性及其硫化氢比色检测
首发时间:2016-04-20
摘要:由于硫化氢(H2S) 在各种生理过程的作用以及它固有的生物毒性,所以发展简单、灵敏的方法检测H2S就显得十分重要。在本文中,我们利用铽-G-四链体-血红素(Tb/G4-hemin)DNA酶作为过氧化物模拟酶提出了在一种检测H2S的比色方法。Tb/G4-heminDNA酶能够催化H2O2氧化 2,2-联氮-二 (3-乙基-苯并噻唑-6-磺酸) 二铵盐(ABTS)产生自由基正离子(ABTSo+)。存在Ag+时,由于破坏了G-四链体的结构,Tb/G4-hemin DNA过氧化物酶的活性被大大地抑制。然而,H2S的加入则通过竞争性结合Ag+,抑制这样的负效应,导致Tb/G4-hemin DNA过氧化物模拟酶活性恢复,最终表现为ABTSo+吸光强度的增加。ABTSo+的吸光强度与浓度范围在20 nM到2 μM 的H2S呈线关系。H2S的检出限为13 nM,这远远低于大多数先前的方法。此外,该方法还具有制备过程简单、重现性好、生物相容性好等特点。
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Colorimetric detection of hydrogen sulfide based on terbium-G-quadruplex-hemin DNAzyme
Abstract:It is of great importance to simply and sensitively detect hydrogen sulfide (H2S) because of its role in various physiological processes as well as its inherent toxicity. In this work, a colorimetric method for H2S detection was developed by employing terbium-G-quadruplex-hemin (Tb/G4-hemin) DNAzyme a peroxidase mimic, which can catalyze the H2O2-mediated oxidation of 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) to produce radical cation (ABTSo+). In the presence of Ag+, the peroxidase-like activityof Tb/G4-hemin DNAzyme can be inhibited significantly owing to the disruption of G-quadruplex structure. However, the addition of H2S can effectively suppress such negative behavior by competitive binding with Ag+, leading to the recovery of the peroxidase-like activity of Tb/G4-hemin DNAzyme, which can be reflected by an increase in absorbance signal of ABTSo+. The absorbance of ABTSo+ was enhanced linearly with increasing the H2S concentration from 20 nM to 2 μM. The detection limit for H2S is 13 nM, which is much lower than most of previous methods. Moreover, the proposed method possesses the features of simple preparation, easy reproducibility and good biocompatibility.
Keywords: Hydrogen sulfide Colorimetric detection G-quadruplex Terbium ion DNAzyme
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铽-G-四链体-血红素复合物的过氧化物氧化酶活性及其硫化氢比色检测
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