Biochemical characterization of a uranyl ion-specific DNAzyme

Andrea K. Brown, Juewen Liu, Ying He, Yi Lu

Research output: Contribution to journalArticlepeer-review


The biochemical characterization of a DNAzyme that is highly specific for uranyl (UO22) ions is described. Sequence alignment, enzyme truncation, and mutation studies have resulted in a conserved sequence that folds into a bulged stem-loop structure. Interestingly, a G-A pair next to the scissile site is important for the uranyl ion-specific DNAzyme; this is reminiscent of the G-T wobble base pair adjacent to the cleavage site that is crucial for the PbII-specific 8-17 DNAzyme activity. Therefore wobble pairs might be important for formation of metal-specific metal-binding sites in DNAzymes. The DNAzyme binds the uranyl ion with a dissociation constant of 469 nm, which is among the strongest metal-binding affinities in nucleic acid enzymes reported to date. This explains why a catalytic beacon fluorescent sensor based on this enzyme has a detection limit (45 pm) that rivals the most-sensitive analytical instrument. It also has over 1 000 000-fold specificity for the uranyl ion over other metal ions. The DNAzyme can carry out multiple turnover reactions that follow the Michaelis -Menten equation, with a kcat of 1.46 min1 and a KM of 463 nm, similar to that of the 8-17 DNAzyme. The pH profile shows a bell-shaped curve that reaches a maximum at pH 5.5, at which the in vitro selection was carried out; this suggests that in vitro selection can be performed to obtain DNAzymes with optimal performance under specific conditions under which practical applications are required. These findings enrich our fundamental understanding of metal-binding sites in nucleic acids and allow the design of sensors with better performance.

Original languageEnglish (US)
Pages (from-to)486-492
Number of pages7
Issue number3
StatePublished - Feb 13 2009


  • Bioinorganic chemistry
  • Biotechnology
  • DNA cleavage
  • DNAzymes
  • Selection
  • Uranyl ion

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology


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