A new method for the detection of ATP using a quantum-dot-tagged aptamer.
Fluorescence resonance energy transfer (FRET) between a quantum dot as donor and an organic fluorophore as acceptor has been widely used for detection of nucleic acids and proteins. In this paper, we developed a new method, characterized by 605-nm quantum dot (605QD) fluorescence intensity increase and corresponding Cy5 fluorescence intensity decrease, to detect adenosine triphosphate (ATP). The new method involved the use of three different oligonucleotides: 3'-biotin-modified DNA that binds to streptavidin-conjugated 605QD; 3'-Cy5-labelled DNA; and a capture DNA consisting of an ATP aptamer and a sequence which could hybridize with both 3'-biotin-modified DNA and 3'-Cy5-labelled DNA. In the absence of the target ATP, the capture DNA binds to 3'-biotin-modified DNA and 3'-Cy5-labelled DNA, bringing quantum dot and Cy5 into close proximity for greater FRET efficiency. When ATP is introduced, the release of the 3'-Cy5-labelled DNA from the hybridization complex took place, triggering 605QD fluorescence intensity increase and corresponding Cy5 fluorescence intensity decrease. Taken together, the virtue of FRET pair 605QD/Cy5 and the property of aptamer-specific conformation change caused by aptamer-ATP interaction, combined with the fluorescence intensity change of both 605QD and Cy5, provide prerequisites for simple and convenient ATP detection.
Chen
Zhang
Z
Department of Microbiology, Hunan Normal University, Changsha, Hunan, 410081, China.
Li
Guang
G
Zhang
Lan
L
Jiang
Junfeng
J
Li
Zhao
Z
Peng
Zhihui
Z
Deng
Le
L
D000255:Adenosine Triphosphate / Q000032:analysis* / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D001710:Biotin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D045663:Quantum Dots; D013194:Staining and Labeling; D019809:Streptavidin / Q000737:chemistry
D052157:Aptamers, Nucleotide; D002232:Carbocyanines; C085321:cyanine dye 5; D001710:Biotin; D000255:Adenosine Triphosphate; D004247:DNA; D019809:Streptavidin