Pyrrolo-dC modified duplex DNA as a novel probe for the sensitive assay of base excision repair enzyme activity

We develop a novel approach to determine formamidopyrimidine DNA glycosylase (Fpg) activity by taking advantage of the unique fluorescence property of pyrrolo-dC (PdC) positioned opposite to 8-oxoguanine (8-oxoG) in duplex DNA. In its initial state, PdC in duplex DNA undergoes the efficient stacking and collisional quenching interactions, showing the low fluorescence signal. In contrast, the presence of Fpg, which specifically removes 8-oxoG and incises resulting apurinic (AP) site, transforms duplex DNA into single-stranded (ss) DNAs. As a result, the intrinsic fluorescence signal of PdC in ssDNA is recovered to exhibit the significantly enhanced fluorescence signal. Based on this Fpg-dependent fluorescence response of PdC, we could reliably determine Fpg activity down to 1.25 U/ml with a linear response from 0 to 50 U/ml. In addition, the diagnostic capability of this strategy was successfully demonstrated by reliably assaying Fpg activity in human blood serum, showing its great potential in the practical applications.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2017-12
Language
English
Keywords

LABEL-FREE; ELECTROCHEMICAL DETECTION; MOLECULAR BEACON; HOGG1 ACTIVITY; FLUORESCENCE; 8-OXOGUANINE; 2-AMINOPURINE; DAMAGE; SUSCEPTIBILITY; NANOCLUSTERS

Citation

BIOSENSORS & BIOELECTRONICS, v.98, pp.210 - 214

ISSN
0956-5663
DOI
10.1016/j.bios.2017.06.052
URI
http://hdl.handle.net/10203/225795
Appears in Collection
CBE-Journal Papers(저널논문)
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