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TitleStructural Basis of Low-pH-Dependent Lysosomal Cholesterol Egress by NPC1 and NPC2.
Journal, issue, pagesCell, Vol. 182, Issue 1, Page 98-111.e18, Year 2020
Publish dateJul 9, 2020
AuthorsHongwu Qian / Xuelan Wu / Ximing Du / Xia Yao / Xin Zhao / Joyce Lee / Hongyuan Yang / Nieng Yan /
PubMed AbstractLysosomal cholesterol egress requires two proteins, NPC1 and NPC2, whose defects are responsible for Niemann-Pick disease type C (NPC). Here, we present systematic structural characterizations that ...Lysosomal cholesterol egress requires two proteins, NPC1 and NPC2, whose defects are responsible for Niemann-Pick disease type C (NPC). Here, we present systematic structural characterizations that reveal the molecular basis for low-pH-dependent cholesterol delivery from NPC2 to the transmembrane (TM) domain of NPC1. At pH 8.0, similar structures of NPC1 were obtained in nanodiscs and in detergent at resolutions of 3.6 Å and 3.0 Å, respectively. A tunnel connecting the N-terminal domain (NTD) and the transmembrane sterol-sensing domain (SSD) was unveiled. At pH 5.5, the NTD exhibits two conformations, suggesting the motion for cholesterol delivery to the tunnel. A putative cholesterol molecule is found at the membrane boundary of the tunnel, and TM2 moves toward formation of a surface pocket on the SSD. Finally, the structure of the NPC1-NPC2 complex at 4.0 Å resolution was obtained at pH 5.5, elucidating the molecular basis for cholesterol handoff from NPC2 to NPC1(NTD).
External linksCell / PubMed:32544384
MethodsEM (single particle)
Resolution3.0 - 4.0 Å
Structure data

21545

6w5r

EMDB-21545, PDB-6w5r:
NPC1 structure in Nanodisc
Method: EM (single particle) / Resolution: 3.6 Å

21546

6w5s

EMDB-21546, PDB-6w5s:
NPC1 structure in GDN micelles at pH 8.0
Method: EM (single particle) / Resolution: 3.0 Å

21547

6w5t

EMDB-21547, PDB-6w5t:
NPC1 structure in GDN micelles at pH 5.5, conformation a
Method: EM (single particle) / Resolution: 3.7 Å

21548

6w5u

EMDB-21548, PDB-6w5u:
NPC1 structure in GDN micelles at pH 5.5, conformation b
Method: EM (single particle) / Resolution: 3.9 Å

21549

6w5v

EMDB-21549, PDB-6w5v:
NPC1-NPC2 complex structure at pH 5.5
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

ChemComp-CLR:
CHOLESTEROL

ChemComp-POV:
(2S)-3-(hexadecanoyloxy)-2-[(9Z)-octadec-9-enoyloxy]propyl 2-(trimethylammonio)ethyl phosphate / phospholipid*YM

Source
  • homo sapiens (human)
KeywordsTRANSPORT PROTEIN / Cholesterol Lysosome

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