A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

被引:38
|
作者
Kermani, Hanie Ahmadzade [1 ]
Hosseini, Morteza [1 ]
Miti, Andrea [2 ,3 ]
Dadmehr, Mehdi [4 ]
Zuccheri, Giampaolo [2 ,3 ]
Hosseinkhani, Saman [5 ]
Ganjali, Mohammad Reza [6 ,7 ]
机构
[1] Univ Tehran, Fac New Sci & Technol, Dept Life Sci Engn, North Kargar St, Tehran 1417466191, Iran
[2] Univ Bologna, Natl Interuniv Consortium Mat Sci & Technol, Dept Pharm & Biotechnol, Nanosci Inst,CNR, Via Irnerio 48, I-40126 Bologna, Italy
[3] Univ Bologna, Natl Interuniv Consortium Mat Sci & Technol, Interdept Ctr Ind Res Hlth Sci & Technol, Nanosci Inst,CNR, Via Irnerio 48, I-40126 Bologna, Italy
[4] Payame Noor Univ, Dept Biotechnol, Lashkarak Rd,POB 19395-4697, Tehran, Iran
[5] Tarbiat Modares Univ, Dept Biochem, POB 14115-111, Tehran, Iran
[6] Univ Tehran, Ctr Excellence Electrochem, Sch Chem, Coll Sci, 16th Azar St,Enghelab Sq,POB 14155-6619, Tehran, Iran
[7] Univ Tehran Med Sci, Endocrinol & Metab Mol Cellular Sci Inst, Biosensor Res Ctr, 10 Jalale Al Ahmad St, Tehran 1411713137, Iran
关键词
Ag/Pt nanoclusters; DNA methyltransferase; Colorimetric detection; Enzyme mimic; CASCADE AMPLIFICATION STRATEGY; HIGHLY SENSITIVE DETECTION; SILVER NANOCLUSTERS; METHYLATION; FLUORESCENCE; BINDING; TARGET; PROBE; RECOGNITION; PLATFORM;
D O I
10.1007/s00216-018-1143-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
DNA methylation catalyzed by DNA methyl transferase (MTase) is a significant epigenetic process for modulating gene expression. Abnormal levels of DNA MTase enzyme have been regarded as a cancer biomarker or a sign of bacterial diseases. We developed a novel colorimetric method to assay M. SssI MTase activity employing peroxidase-like activity of DNA template Ag/Pt NCs without using restriction enzymes. Based on inhibiting the peroxidase reaction that occurred in the TMB-H2O2 system, in the presence of MTase, a highly sensitive and selective colorimetric biosensor was fabricated with a detection limit (LOD) of 0.05 U/mL and a linear range from 0.5 to 10 U/mL. The changes in absorption intensity were monitored to quantify the M. SssI activity. This strategy had a high selectivity over other proteins. Furthermore, it is also demonstrated that this method can be used for the evaluation and screening of inhibitors for DNA MTase.
引用
收藏
页码:4943 / 4952
页数:10
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