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.
Department of Microbiology, Hunan Normal University, Changsha, Hunan, 410081, China.
Zhang
Z
Chen
Guang
G
Li
Lan
L
Zhang
Junfeng
J
Jiang
Zhao
Z
Li
Zhihui
Z
Peng
Le
L
Deng
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