Cover Image

Structural and biochemical analysis of ligand binding in yeast Niemann-Pick type C1-related protein

In eukaryotes, integration of sterols into the vacuolar/lysosomal membrane is critically dependent on the Niemann-Pick type C (NPC) system. The system consists of an integral membrane protein, called NCR1 in yeast, and NPC2, a luminal soluble protein that transfers sterols to the N-terminal domain (...

Full description

Saved in:
Bibliographic Details
Main Authors: Nel, Lynette (Author) , Thaysen, Katja (Author) , Jamecna, Denisa (Author) , Olesen, Esben (Author) , Szomek, Maria (Author) , Langer, Julia (Author) , Frain, Kelly M. (Author) , Höglinger, Doris (Author) , Wüstner, Daniel (Author) , Pedersen, Bjørn P. (Author)
Format: Article (Journal)
Language:English
Published: 2025
In: Life science alliance
Year: 2025, Volume: 8, Issue: 1, Pages: 1-15
ISSN:2575-1077
DOI:10.26508/lsa.202402990
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.26508/lsa.202402990
Verlag, kostenfrei, Volltext: https://www.life-science-alliance.org/content/8/1/e202402990
Get full text
Author Notes:Lynette Nel, Katja Thaysen, Denisa Jamecna, Esben Olesen, Maria Szomek, Julia Langer, Kelly M. Frain, Doris Höglinger, Daniel Wüstner, Bjørn P. Pedersen
Description
Summary:In eukaryotes, integration of sterols into the vacuolar/lysosomal membrane is critically dependent on the Niemann-Pick type C (NPC) system. The system consists of an integral membrane protein, called NCR1 in yeast, and NPC2, a luminal soluble protein that transfers sterols to the N-terminal domain (NTD) of NCR1 before membrane integration. Both proteins have been implicated in sterol homeostasis of yeast and humans. Here, we investigate sterol and lipid binding of the NCR1/NPC2 transport system and determine crystal structures of the sterol binding NTD. The NTD binds both ergosterol and cholesterol, with nearly identical conformations of the binding pocket. Apart from sterols, the NTD can also bind fluorescent analogs of phosphatidylinositol, phosphatidylcholine, and phosphatidylserine, as well as sphingosine and ceramide. We confirm the multi-lipid scope of the NCR1/NPC2 system using photo-crosslinkable and clickable lipid analogs, namely, pac-cholesterol, pac-sphingosine, and pac-ceramide. Finally, we reconstitute the transfer of pac-sphingosine from NPC2 to the NTD in vitro. Collectively, our results support that the yeast NPC system can work as versatile machinery for vacuolar homeostasis of structurally diverse lipids, besides ergosterol. - Graphical Abstract - <img class="highwire-fragment fragment-image" alt="Figure" src="https://www.life-science-alliance.org/content/lsa/8/1/e202402990/F1.medium.gif" width="440" height="218"/>Download figureOpen in new tabDownload PowerPoint
Item Description:Veröffentlicht: 25 October 2024
Gesehen am 04.04.2025
Physical Description:Online Resource
ISSN:2575-1077
DOI:10.26508/lsa.202402990

Similar Items