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- Title
Identification of NPC1 as the target of U18666A, an inhibitor of lysosomal cholesterol export and Ebola infection.
- Authors
Feiran Lu; Qiren Liang; Abi-Mosleh, Lina; Das, Akash; De Brabander, Jef K.; Goldstein, Joseph L.; Brown, Michael S.
- Abstract
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.
- Subjects
EBOLA virus disease; NIEMANN-Pick diseases; PHYSIOLOGICAL effects of cholesterol; POINT mutation (Biology); VIRAL mutation; CROSSLINKING (Polymerization)
- Publication
eLife, 2015, p1
- ISSN
2050-084X
- Publication type
Article
- DOI
10.7554/eLife.12177