A new method for the in situ assay of α-glucosidase activity and inhibitor screening through an enzyme substrate-mediated DNA hybridization chain reaction
| Autor: | Junfeng Liu, Xiquan Yue, Xu Chen, Lihong Su, Longpo Zheng, Juan Zhang |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
In situ
chemistry.chemical_classification biology Chemistry DNA–DNA hybridization Substrate (chemistry) 02 engineering and technology General Chemistry 010402 general chemistry 021001 nanoscience & nanotechnology 01 natural sciences Combinatorial chemistry Catalysis Enzyme assay 0104 chemical sciences Hydrolysis Enzyme Materials Chemistry biology.protein 0210 nano-technology human activities Chain reaction |
| Zdroj: | New Journal of Chemistry. 43:9458-9465 |
| ISSN: | 1369-9261 1144-0546 |
| Popis: | In this work, a new strategy has been proposed for the in situ analysis of α-glucosidase (α-Glu) based on the small molecule-mediated DNA hybridization chain reaction (HCR). As an important digestion enzyme, α-Glu could catalyze the hydrolysis of the substrate para-aminophenyl-α-D-glucopyranoside (pAPG) into para-aminophenyl (pAP) and α-D-glucopyranoside. Among them, pAPG with cis-diols groups could coordinate with the boronic acid-tagged initiation chain to drive HCR around the assembled magnetic nanospheres (AMNSs). Nevertheless, the product of the catalytic reaction pAP did not have the capability to capture the boronic acid-tagged initiation chain onto the surface of AMNSs, due to which HCR did not occur. Thus, the α-Glu activity was correlated with the extent of HCR around AMNSs and it could be tested in situ in a cell medium with the aid of a magnet. The enzyme activity was analyzed over a wide linear range from 0.4 U mL−1 to 1.7 U mL−1 with the lowest detection limit of 0.023 U mL−1. Furthermore, the established method could also be successfully utilized for the in situ evaluation of enzyme inhibitors including gallic acid and quercetin. This study may provide a new insight for the development of a new method for the in situ analysis of α-Glu in a cell medium. |
| Databáze: | OpenAIRE |
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