Simple and real-time colorimetric assay for glycosidases activity using functionalized gold nanoparticles and its application for inhibitor screening.
Simple and real-time colorimetric assay for glycosidases activity using functionalized gold nanoparticles and its application for inhibitor screening.
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DOI：
10.1021/ac301677v
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发表时间：
2012-10
影响因子：
7.4
通讯作者：
Zhanghua Zeng;S. Mizukami;K. Kikuchi
中科院分区：
文献类型：
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作者：
Zhanghua Zeng;S. Mizukami;K. Kikuchi
近年来，酶的实时检测在生物医学研究中受到了极大的关注。自焚消除是对外界刺激的反应，通过一系列消除过程，将一个多组分的结构自发地和不可逆地分解成其组成片段。在这里，我们报告了一种简单而实时的糖苷酶(β-半乳糖苷酶和β-葡萄糖苷酶)比色法。在功能化的金纳米颗粒(Gal-Lip-AuNPs和GLC-Lip-AuNPs，其中AuNPs表示金纳米颗粒)上显示的酶裂解触发器后，利用自焚消除来释放胺引起聚集和颜色变化。β-半乳糖苷酶和β-葡萄糖苷酶的检测下限在20min时分别为9.2nM和22.3nM，并随着时间的推移略有提高。因此，成功地实时检测了糖苷酶活性，该技术可用于基于实时比色变化的糖苷酶抑制剂筛选。
The development of real-time assays for enzymes has been receiving a great deal of attention in biomedical research recently. Self-immolative elimination is the spontaneous and irreversible disassembly of a multicomponent construct into its constituent fragments through a cascade of elimination processes, in response to external stimuli. Here, we report a simple and real-time colorimetric assay for glycosidases (β-galactosidase and β-glucosidase). Self-immolative elimination was utilized to release amines to give rise to aggregation and color change by electrostatic attraction after cleavage of the trigger by enzymes displayed on functionalized gold nanoparticles (Gal-Lip-AuNPs and Glc-Lip-AuNPs, where AuNPs denotes gold nanoparticles). The detection limits for β-galactosidase and β-glucosidase were as low as 9.2 and 22.3 nM at 20 min, and they improved slightly over time. Thus, glycosidase activity was detected successfully in real time, and this technique could be used for glycosidase inhibitor screening, based on real-time colorimetric variation.