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

Lynette Nel, Katja Thaysen, Denisa Jamecna, Esben Olesen, Julia Langer, Kelly May Frain, Maria Szomek, Doris Höglinger, Daniel Wüstner*, Bjørn Panyella Pedersen*

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Abstract

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 pacceramide.
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.
OriginalsprogEngelsk
Artikelnummere202402990
TidsskriftLife Science Alliance
Vol/bind8
Nummer1
Antal sider15
ISSN2575-1077
DOI
StatusUdgivet - jan. 2025

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