Reduction-triggered fluorescence probe for peptide-templated reactions.
We developed a new nucleic acid-based fluorescence probe for protein detection. The method is based on the scission of an aptamer into two probes, which are then attached with a chemically reactive fluorogenic compound. The protein-dependent association of the two probes accelerates a reduction-triggered fluorogenic reaction and indicates the presence of the target protein, which is detected using a fluorescence readout. The fluorescence signal is generated via the deprotection of the azidomethyl group of fluorescein. The arginine-rich motif peptide of the human immunodeficiency virus-1 Rev protein was targeted by this type of probe. Emission was detected at 522 nm and was enhanced by about 19.4-fold in the presence of the target peptide. An oligonucleotide-based reduction-triggered fluorescence probe was successfully applied to the detection of the Rev peptide in solution.
Shibata
Aya
A
Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Abe
Hiroshi
H
Furukawa
Kazuhiro
K
Tsuneda
Satoshi
S
Ito
Yoshihiro
Y
D020816:Amino Acid Motifs; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D009696:Nucleic Acids / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D010084:Oxidation-Reduction; D010455:Peptides / Q000032:analysis*; D013997:Time Factors; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000737:chemistry
D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D009841:Oligonucleotides; D010455:Peptides; D054321:rev Gene Products, Human Immunodeficiency Virus