title: A label-free fluorescence strategy for sensitive detection of ATP based on the ligation-triggered super-sandwich.
abstract: In this study, a label-free fluorescence strategy for sensitive detection of ATP based on the ligation-triggered super-sandwich is reported. We designed a double-stranded DNA (ds-DNA) probe as the substrate of ATP-dependent ligation. SYBR Green I (SG I), a double-duplex DNA specific dye, was employed as the readout signal. In the absence of ATP, the ligation would not occur and the ds-DNA remained intact. Further, a weak fluorescence could be observed due to the intercalation of SG I into the grooves of the ds-DNA probe. In the presence of ATP, T4 DNA ligase would catalyse the ligation between 3'-OH and 5'-PO4 ends between ds-DNA probes. As a result, more binding sites of the SG I were generated and a fluorescence enhancement was obtained. This method showed a good sensitivity with a detection limit of 200 pM and could perfectly discriminate ATP from its analogs.
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Chunshui
authors.initials: C
authors.lastname: Lin
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Yiying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Zhixiong
authors.initials: Z
authors.lastname: Cai
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Yaqi
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China; State Key Laboratory of Physical Chemistry of Solid Surfaces, The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China.
authors.forename: Chaoyong James
authors.initials: CJ
authors.lastname: Yang
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China; State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China. Electronic address: xichen@xmu.edu.cn.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D015342:DNA Probes; D000255:Adenosine Triphosphate
keywords: ATP; Fluorescence; Label-free; SYBR Green I; Super-sandwich; T4 DNA ligase