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.
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated
