Identification of NPC1 as the target of U18666A, an inhibitor of lysosomal cholesterol export and Ebola infection
| العنوان: | Identification of NPC1 as the target of U18666A, an inhibitor of lysosomal cholesterol export and Ebola infection |
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| المؤلفون: | Jef K. De Brabander, Joseph L. Goldstein, Feiran Lu, Michael S. Brown, Akash Das, Lina Abi-Mosleh, Qiren Liang |
| المصدر: | eLife, Vol 4 (2015) eLife |
| بيانات النشر: | eLife Sciences Publications Ltd, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | medicine.disease_cause, Biochemistry, chemistry.chemical_compound, U18666A, Niemann-Pick C1, crosslinking, Enzyme Inhibitors, Biology (General), Membrane Glycoproteins, biology, General Neuroscience, Chinese hamster ovary cell, Anticholesteremic Agents, Intracellular Signaling Peptides and Proteins, General Medicine, Ebolavirus, 3. Good health, Cell biology, Cholesterol, Medicine, Androstenes, lipids (amino acids, peptides, and proteins), Cricetulus, Research Article, Human, congenital, hereditary, and neonatal diseases and abnormalities, QH301-705.5, Science, CHO Cells, Antiviral Agents, General Biochemistry, Genetics and Molecular Biology, Viral entry, Niemann-Pick C1 Protein, cholesterol transport, medicine, Animals, Ebola virus, General Immunology and Microbiology, Point mutation, endosomes/lysosomes, Cell Biology, Virus Internalization, biology.organism_classification, Sterol, sterol-sensing domain, chemistry, Other, NPC1, Carrier Proteins |
| الوصف: | Niemann-Pick C1 (NPC1) is a lysosomal membrane protein that exports cholesterol derived from receptor-mediated uptake of LDL, and it also mediates cellular entry of Ebola virus. Cholesterol export is inhibited by nanomolar concentrations of U18666A, a cationic sterol. To identify the target of U18666A, we synthesized U-X, a U18666A derivative with a benzophenone that permits ultraviolet-induced crosslinking. When added to CHO cells, U-X crosslinked to NPC1. Crosslinking was blocked by U18666A derivatives that block cholesterol export, but not derivatives lacking blocking activity. Crosslinking was prevented by point mutation in the sterol-sensing domain (SSD) of NPC1, but not by point mutation in the N-terminal domain (NTD). These data suggest that the SSD contains a U18666A-inhibitable site required for cholesterol export distinct from the cholesterol-binding site in the NTD. Inasmuch as inhibition of Ebola requires 100-fold higher concentrations of U18666A, the high affinity U16888A-binding site is likely not required for virus entry. DOI: http://dx.doi.org/10.7554/eLife.12177.001 eLife digest Cholesterol is a type of fat molecule and is a vital component of animal cell membranes. It is taken up into cells within particles called low density lipoproteins (LDLs) that are then digested in cell compartments known as lysosomes to release the cholesterol. Then, the cholesterol leaves the lysosome with the help of a transport protein called NPC1. Mutations in the gene that encodes NPC1 lead to the accumulation of cholesterol in lysosomes; this can cause a devastating illness that affects the brain, liver and other organs. The NPC1 protein also plays a crucial role in allowing Ebola viruses to infect animal cells and multiply. U18666A is a drug that blocks the movement of cholesterol out of lysosomes and also inhibits Ebola virus infections, but it was not known what components it targets in cells. Lu et al. used a technique called ultraviolet-induced crosslinking to identify the proteins that U18666A can bind to. The experiments show that U18666A can directly bind to a site that is within a section of the NPC1 protein called the sterol-sensing domain. The binding of U18666A to this site blocks the movement of cholesterol out of lysosomes. Lu et al.’s findings indicate that the sterol-sensing domain of NPC1 plays a crucial role in cholesterol’s export from lysosomes. A future challenge is to use structural biology techniques (such as X-ray crystallography or cryo-electron microscope tomography) to understand the three-dimensional structure of NPC1. DOI: http://dx.doi.org/10.7554/eLife.12177.002 |
| اللغة: | English |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7846400e7141f2c3663993bdbdfdfa07 https://elifesciences.org/articles/12177 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....7846400e7141f2c3663993bdbdfdfa07 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |