Real-time monitoring of disintegration activity of catalytic core domain of HIV-1 integrase using molecular beacon

Anal Biochem. 2013 Sep 15;440(2):120-2. doi: 10.1016/j.ab.2013.05.032. Epub 2013 Jun 6.

Abstract

HIV-1 integrase, an essential enzyme for retroviral replication, is a validated target for anti-HIV therapy development. The catalytic core domain of integrase (IN-CCD) is capable of catalyzing disintegration reaction. In this work, a hairpin-shaped disintegration substrate was designed and validated by enzyme-linked immunosorbent assay; a molecular beacon-based assay was developed for disintegration reaction of IN-CCD. Results showed that the disintegration substrate could be recognized and catalyzed by IN-CCD, and the disintegration reaction can be monitored according to the increase of fluorescent signal. The assay can be applied to real-time detection of disintegration with advantages of simplicity, high sensitivity, and excellent specificity.

Keywords: Disintegration; HIV-1; IN-CCD; Integrase; Molecular beacon.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Catalytic Domain*
  • Enzyme Assays / methods*
  • HIV Integrase / chemistry*
  • HIV Integrase / metabolism*
  • HIV-1 / enzymology*
  • Oligonucleotide Probes / genetics
  • Oligonucleotide Probes / metabolism*
  • Time Factors

Substances

  • Oligonucleotide Probes
  • HIV Integrase