Publikation: Protein-dependent ribozymes report molecular interactions in real time
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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.
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HARTIG, Jörg S., S. Hani NAJAFI-SHOUSHTARI, Imke GRÜNE, Amy YAN, Andrew D. ELLINGTON, Michael FAMULOK, 2002. Protein-dependent ribozymes report molecular interactions in real time. In: Nature Biotechnology. 2002, 20(7), pp. 717-722. ISSN 1087-0156. Available under: doi: 10.1038/nbt0702-717BibTex
@article{Hartig2002-07Prote-15661, year={2002}, doi={10.1038/nbt0702-717}, title={Protein-dependent ribozymes report molecular interactions in real time}, number={7}, volume={20}, issn={1087-0156}, journal={Nature Biotechnology}, pages={717--722}, author={Hartig, Jörg S. and Najafi-Shoushtari, S. Hani and Grüne, Imke and Yan, Amy and Ellington, Andrew D. and Famulok, Michael} }
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