Protein-dependent ribozymes report molecular interactions in real time. Academic Article uri icon

Overview

abstract

  • Most approaches to monitoring interactions between biological macromolecules require large amounts of material, rely upon the covalent modification of an interaction partner, or are not amenable to real-time detection. We have developed a generalizable assay system based on interactions between proteins and reporter ribozymes. The assay can be configured in a modular fashion to monitor the presence and concentration of a protein or of molecules that modulate protein function. We report two applications of the assay: screening for a small molecule that disrupts protein binding to its nucleic acid target and screening for protein protein interactions. We screened a structurally diverse library of antibiotics for small molecules that modulate the activity of HIV-1 Rev-responsive ribozymes by binding to Rev. We identified an inhibitor that subsequently inhibited HIV-1 replication in cells. A simple format switch allowed reliable monitoring of domain-specific interactions between the blood-clotting factor thrombin and its protein partners. The rapid identification of interactions between proteins or of compounds that disrupt such interactions should have substantial utility for the drug-discovery process.

publication date

  • July 1, 2002

Research

keywords

  • DNA, Single-Stranded
  • Gene Products, rev
  • Protein Interaction Mapping
  • RNA, Catalytic

Identity

Scopus Document Identifier

  • 0035989643

PubMed ID

  • 12089558

Additional Document Info

volume

  • 20

issue

  • 7