Novel Class of Potential Therapeutics that Target Ricin Retrograde Translocation
| Autor: | Zerlina Lau, Veronika Redmann, Thomas J. Gardner, Keita Morohashi, Dan P. Felsenfeld, Domenico Tortorella |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Proteasome Endopeptidase Complex
Health Toxicology and Mutagenesis Green Fluorescent Proteins lcsh:Medicine Ricin Biology Endoplasmic Reticulum Toxicology Article Green fluorescent protein Small Molecule Libraries Cell membrane 03 medical and health sciences chemistry.chemical_compound Cell Line Tumor medicine Humans Toxins Biological 030304 developmental biology 0303 health sciences dislocation Ricinus Endoplasmic reticulum Ribosome-inactivating protein Cell Membrane 030302 biochemistry & molecular biology lcsh:R small molecule inhibitors high-content screen stabilization Cytosol HEK293 Cells medicine.anatomical_structure retrograde translocation chemistry Biochemistry egfp Protein Biosynthesis ricin toxin Proteasome inhibitor ribosome-inactivating protein Intracellular medicine.drug |
| Zdroj: | Toxins, Vol 6, Iss 1, Pp 33-53 (2013) Toxins Volume 6 Issue 1 Pages: 33-53 |
| ISSN: | 2072-6651 |
| Popis: | Ricin toxin, an A-B toxin from Ricinus communis, induces cell death through the inhibition of protein synthesis. The toxin binds to the cell surface via its B chain (RTB) followed by its retrograde trafficking through intracellular compartments to the ER where the A chain (RTA) is transported across the membrane and into the cytosol. Ricin A chain is transported across the ER membrane utilizing cellular proteins involved in the disposal of aberrant ER proteins by a process referred to as retrograde translocation. Given the current lack of therapeutics against ricin intoxication, we developed a high-content screen using an enzymatically attenuated RTA chimera engineered with a carboxy-terminal enhanced green fluorescent protein (RTA(E177Q)egfp) to identify compounds that target RTA retrograde translocation. Stabilizing RTA(E177Q)egfp through the inclusion of proteasome inhibitor produced fluorescent peri-nuclear granules. Quantitative analysis of the fluorescent granules provided the basis to discover compounds from a small chemical library (2080 compounds) with known bioactive properties. Strikingly, the screen found compounds that stabilized RTA molecules within the cell and several compounds limited the ability of wild type RTA to suppress protein synthesis. Collectively, a robust high-content screen was developed to discover novel compounds that stabilize intracellular ricin and limit ricin intoxication. |
| Databáze: | OpenAIRE |
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