Triazoles inhibit cholesterol export from lysosomes by binding to NPC1
| Autor: | Joseph L. Goldstein, Qiren Liang, Jef K. De Brabander, Xiaochun Li, Akash Das, Feiran Lu, Michael S. Brown, Michael N. Trinh |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
congenital hereditary and neonatal diseases and abnormalities Antifungal Agents Antifungal drug Triazole CHO Cells Biology Endocytosis Cell Line 03 medical and health sciences chemistry.chemical_compound Cricetulus 0302 clinical medicine Protein Domains hemic and lymphatic diseases medicine Animals Binding site Lipid bilayer Binding Sites Membrane Glycoproteins Multidisciplinary Cholesterol nutritional and metabolic diseases Biological Transport Biological Sciences Triazoles Ketoconazole 030104 developmental biology chemistry Biochemistry Androstenes lipids (amino acids peptides and proteins) Itraconazole NPC1 Carrier Proteins Lysosomes 030217 neurology & neurosurgery Protein Binding medicine.drug |
| Zdroj: | Proceedings of the National Academy of Sciences. 114:89-94 |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | Niemann–Pick C1 (NPC1), a membrane protein of lysosomes, is required for the export of cholesterol derived from receptor-mediated endocytosis of LDL. Lysosomal cholesterol export is reportedly inhibited by itraconazole, a triazole that is used as an antifungal drug [Xu et al. (2010) Proc Natl Acad Sci USA 107:4764–4769]. Here we show that posaconazole, another triazole, also blocks cholesterol export from lysosomes. We prepared P-X, a photoactivatable cross-linking derivative of posaconazole. P-X cross-linked to NPC1 when added to intact cells. Cross-linking was inhibited by itraconazole but not by ketoconazole, an imidazole that does not block cholesterol export. Cross-linking of P-X was also blocked by U18666A, a compound that has been shown to bind to NPC1 and inhibit cholesterol export. P-X also cross-linked to purified NPC1 that was incorporated into lipid bilayer nanodiscs. In this in vitro system, cross-linking of P-X was inhibited by itraconazole, but not by U18666A. P-X cross-linking was not prevented by deletion of the N-terminal domain of NPC1, which contains the initial binding site for cholesterol. In contrast, P-X cross-linking was reduced when NPC1 contained a point mutation (P691S) in its putative sterol-sensing domain. We hypothesize that the sterol-sensing domain has a binding site that can accommodate structurally different ligands. |
| Databáze: | OpenAIRE |
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