Selection of DNA aptamers that inhibit enzymatic activity of PQQGDH and its application.
We screened DNA aptamers against the water-soluble quinoprotein glucose dehydrogenase (PQQGDH) with a competitive selection method. PQQGDH is a dimeric enzyme that consists of 50-kDa subunits, and has high catalytic activity for glucose (about 5000 U/mg), therefore is used for glucose sensors in the market. PQQGDH is an excellent molecular recognition device for biosensor for diagnosis. The SELEX was performed using a competitive selection method which enables us to select aptamer having high specificity for a target molecule. After 6 rounds screening, we obtained 2 aptamers which bind to PQQGDH with high affinity and specificity. One aptamer inhibited PQQGDH activity. In addition, we constructed a dimer aptamer linked by linker sequences expecting high affinity compared to the monomer aptamer. As the result of Aptamer Blotting, the dimer aptamer showed higher affinity than the monomer.
Ikebukuro
Kazunori
K
Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan. ikebu@cc.tuat.ac.jp
Takase
Madoka
M
Sode
Koji
K
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques; D004277:DNA, Single-Stranded / Q000378:metabolism; D019281:Dimerization; D005948:Glucose Dehydrogenases / Q000378:metabolism*; D015336:Molecular Probe Techniques; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)