DNA Detection and Signal Amplification via an Engineered Allosteric Enzyme

• Published in: Journal of the American Chemical Society Vol. 125; no. 2; pp. 344 - 345
• Main Authors: Saghatelian, Alan, Guckian, Kevin M, Thayer, Desiree A, Ghadiri, M. Reza
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15.01.2003
• Subjects: Bacillus cereus - enzymology; Bacillus cereus - genetics; Biological and medical sciences; DNA - analysis; DNA, Single-Stranded - chemistry; Fundamental and applied biological sciences. Psychology; Kinetics; Metalloendopeptidases - chemistry; Metalloendopeptidases - genetics; Metalloendopeptidases - metabolism; Miscellaneous; Molecular and cellular biology; Molecular genetics; Oligopeptides - chemistry; Oligopeptides - metabolism; Protease Inhibitors - chemistry; Signal Processing, Computer-Assisted; Amplification; Regulation(control); Allosteric regulation; DNA; Inhibition; Detection; Signal analysis
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja027885u

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.