Disulfide-Trapping Identifies a New, Effective Chemical Probe for Activating the Nuclear Receptor Human LRH-1 (NR5A2)
| Autor: | Elena P. Sablin, Miyuki Suzawa, John M. Bruning, Pamela M. England, Sam Irvy, Holly A. Ingraham, Felipe de Jesus Cortez, Matthew P. Jacobson, Robert J. Fletterick |
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| Přispěvatelé: | Lodola, Alessio |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cytoplasmic and Nuclear Carboxylic Acids lcsh:Medicine Receptors Cytoplasmic and Nuclear Ligands Biochemistry chemistry.chemical_compound Binding Analysis Receptors Small interfering RNAs Disulfides Amino Acids Receptor lcsh:Science Phospholipids Multidisciplinary Chemistry Organic Compounds Liver Disease Hep G2 Cells Lipids Nucleic acids Molecular Docking Simulation 5.1 Pharmaceuticals Physical Sciences Development of treatments and therapeutic interventions Lead compound Cell Binding Assay Hydrophobic and Hydrophilic Interactions Biotechnology Research Article Agonist medicine.drug_class General Science & Technology Library Screening Research and Analysis Methods 03 medical and health sciences Thiols In vivo medicine Genetics Sulfur Containing Amino Acids Humans Cysteine Molecular Biology Techniques Non-coding RNA Molecular Biology Chemical Characterization Virtual screening Molecular Biology Assays and Analysis Techniques Liver receptor homolog-1 lcsh:R Organic Chemistry Chemical Compounds Biology and Life Sciences Proteins Amides Gene regulation 030104 developmental biology Nuclear receptor Molecular Probes RNA lcsh:Q Gene expression Digestive Diseases Acids |
| Zdroj: | PLoS ONE Cortez, FDJ; Suzawa, M; Irvy, S; Bruning, JM; Sablin, E; Jacobson, MP; et al.(2016). Disulfide-Trapping Identifies a New, Effective Chemical Probe for Activating the Nuclear Receptor Human LRH-1 (NR5A2). PLOS ONE, 11(7). doi: 10.1371/journal.pone.0159316. UC San Francisco: Retrieved from: http://www.escholarship.org/uc/item/2bp084k5 PloS one, vol 11, iss 7 PLoS ONE, Vol 11, Iss 7, p e0159316 (2016) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0159316. |
| Popis: | Conventional efforts relying on high-throughput physical and virtual screening of large compound libraries have failed to yield high-efficiency chemical probes for many of the 48 human nuclear receptors. Here, we investigated whether disulfide-trapping, an approach new to nuclear receptors, would provide effective lead compounds targeting human liver receptor homolog 1 (hLRH-1, NR5A2). Despite the fact that hLRH-1 contains a large ligand binding pocket and binds phospholipids with high affinity, existing synthetic hLRH-1 ligands are of limited utility due to poor solubility, low efficacy or significant off-target effects. Using disulfide-trapping, we identified a lead compound that conjugates with remarkably high-efficiency to a native cysteine residue (Cys346) lining the hydrophobic cavity in the ligand binding domain of hLRH-1. Guided by computational modeling and cellular assays, the lead compound was elaborated into ligands PME8 and PME9 that bind hLRH-1 reversibly (no cysteine reactivity) and increase hLRH-1 activity in cells. When compared with the existing hLRH-1 synthetic agonist RJW100, both PME8 and PME9 showed comparable induction of the LRH-1 dependent target gene CYP24A1 in human HepG2 cells, beginning as early as 3 h after drug treatment. The induction is specific as siRNA-mediated knock-down of hLRH-1 renders both PME8 and PME9 ineffective. These data show that PME8 and PME9 are potent activators of hLRH-1 and suggest that with further development this lead series may yield useful chemical probes for manipulating LRH-1 activity in vivo. |
| Databáze: | OpenAIRE |
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