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PMID: 18823126 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of sterol transport between membranes and NPC2.

Biochemistry ·Vol. 47 ·No. 42 ·2008-10-21 ·Pages 11134-43

Xu Z, Farver W, Kodukula S, Storch J

Abstract

Niemann-Pick disease type C (NPC) is caused by defects in either the NPC1 or NPC2 gene and is characterized by accumulation of cholesterol and glycolipids in the late endosome/lysosome compartment. NPC2 is an intralysosomal protein that binds cholesterol in vitro. Previous studies demonstrated rapid rates of cholesterol transfer from NPC2 to model membranes [Cheruku, S. R., et al. (2006) J. Biol. Chem. 281, 31594-31604]. To model the potential role of NPC2 as a lysosomal cholesterol export protein, in this study we used fluorescence spectroscopic approaches to examine cholesterol transfer from membranes to NPC2, assessing the rate, mechanism, and regulation of this transport step. In addition, we examined the effect of NPC2 on the rate and kinetic mechanism of intermembrane sterol transport, to model the movement of cholesterol from internal lysosomal membranes to the limiting lysosomal membrane. The results support the hypothesis that NPC2 plays an important role in endo/lysosomal cholesterol trafficking by markedly accelerating the rates of cholesterol transport. Rates of sterol transfer from and between membranes were increased by as much as 2 orders of magnitude by NPC2. The transfer studies indicate that the mechanism of NPC2 action involves direct interaction of the protein with membranes. Such interactions were observed directly using FTIR spectroscopy and protein tryptophan spectral shifts. Additionally, cholesterol transfer by NPC2 was found to be greatly enhanced by the unique lysosomal phospholipid lyso-bisphosphatidic acid (LBPA), suggesting an important role for LBPA in NPC2-mediated cholesterol trafficking.

MeSH Terms
Animals Biological Transport, Active/drug effects Carrier Proteins/chemistry,genetics,metabolism Cattle Cholesterol/metabolism Dolichols/pharmacology Endosomes/metabolism Ergosterol/analogs & derivatives,metabolism Fluorescent Dyes Gangliosides/pharmacology Glycoproteins/chemistry,genetics,metabolism Humans In Vitro Techniques Intracellular Membranes/metabolism Kinetics Lysosomes/metabolism Niemann-Pick Disease, Type C/genetics,metabolism Recombinant Proteins/chemistry,genetics,metabolism Spectrometry, Fluorescence Spectroscopy, Fourier Transform Infrared Sterols/metabolism Tryptophan/chemistry Vesicular Transport Proteins/chemistry,genetics,metabolism
Chemicals
Carrier Proteins Dolichols Fluorescent Dyes Gangliosides Glycoproteins NPC2 protein, human Recombinant Proteins Sterols Vesicular Transport Proteins dehydroergosterol Tryptophan Cholesterol Ergosterol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Xu Zhi
Department of Nutritional Sciences, School of Environmental and Biological Sciences, Rutgers, the State University of New Jersey, New Brunswick, New Jersey 08901, USA.
Farver William
Kodukula Sarala
Storch Judith
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Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
1520-4995
DOI
10.1021/bi801328u
Published
2008-10-21
Epub
2008-00-30
Pages
11134-43
Language
English
Region
United States
NLM ID
0370623
PMCID
PMC4355403
Subset
IM
Grants
NIDDK NIH HHS · R01 DK038389 · United States
External Links
PubMed source DOI: 10.1021/bi801328u PMC Full Text View journal details
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