title: Selection of DNA aptamers that inhibit enzymatic activity of PQQGDH and its application.
abstract: 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.
authors.affiliation: 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
authors.forename: Kazunori
authors.initials: K
authors.lastname: Ikebukuro
authors.forename: Madoka
authors.initials: M
authors.lastname: Takase
authors.forename: Koji
authors.initials: K
authors.lastname: Sode
mesh_terms: 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
chemical_list: D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)