DNA detection and signal amplification via an engineered allosteric enzyme

J Am Chem Soc. 2003 Jan 15;125(2):344-5. doi: 10.1021/ja027885u.

Abstract

Rapid, sensitive, and sequence-specific DNA detection can be achieved in one step using an engineered intrasterically regulated enzyme. The semi-synthetic inhibitor-DNA-enzyme (IDE) construct (left) rests in the inactive state but upon exposure to a complementary DNA sequence undergoes a DNA hybridization-triggered allosteric enzyme activation (right). The ensuing rapid substrate turnover provides the built-in signal amplification mechanism for detecting approximately 10 fmol DNA in less than 3 min under physiological conditions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacillus cereus / enzymology*
  • Bacillus cereus / genetics
  • DNA / analysis*
  • DNA, Single-Stranded / chemistry
  • Kinetics
  • Metalloendopeptidases / chemistry*
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism
  • Oligopeptides / chemistry
  • Oligopeptides / metabolism
  • Protease Inhibitors / chemistry*
  • Signal Processing, Computer-Assisted

Substances

  • DNA, Single-Stranded
  • Oligopeptides
  • Protease Inhibitors
  • DNA
  • Metalloendopeptidases