The Niemann-Pick C1 and caveolin-1 proteins interact to modulate efflux of low density lipoprotein-derived cholesterol from late endocytic compartments

David Jelinek, Randy A Heidenreich, Robert A Orlando, William S Garver


The Niemann-Pick C1 (NPC1) protein has a central role in regulating the efflux of lipoprotein-derived cholesterol from late endosomes/lysosomes and transport to other cellular compartments. Since the NPC1 protein has been shown to regulate the transport of cholesterol to cellular compartments enriched with the ubiquitous cholesterol-binding and transport protein caveolin-1, the present study was performed to determine whether the NPC1 and caveolin-1 proteins interact and function to modulate efflux of low density lipoprotein (LDL)-derived cholesterol from endocytic compartments. To perform these studies, normal human fibroblasts were grown in media with lipoprotein-deficient serum (LPDS) or media with LPDS supplemented with purified human LDL. The results indicated reciprocal co-immunoprecipitation and partial co-localization of the NPC1 and caveolin-1 proteins that was decreased when fibroblasts were grown in media with LDL. Consistent with interaction of the NPC1 and caveolin-1 proteins, a highly conserved caveolin-binding motif was identified within a cytoplasmic loop located adjacent to the sterol-sensing domain (SSD) of the NPC1 protein. To examine the functional relevance of this interaction, fibroblasts were transfected with caveolin-1 siRNA and found to accumulate increased amounts of LDL-derived cholesterol within late endosomes/lysosomes. Together, this report presents novel results demonstrating that the NPC1 and caveolin-1 proteins interact to modulate efflux of LDL-derived cholesterol from late endocytic compartments.


caveolin-1; cholesterol; endosomes; Niemann-Pick C1


Bist A, Fielding PE & Fielding CJ 1997 Two sterol regulatory element-like sequences mediate up-regulation of caveolin gene transcription in response to low density lipoprotein free cholesterol. Proc Natl Acad Sci USA 94 10693-10698

Blanchette-Mackie EJ, Dwyer NK, Amende LM, Kruth HS, Butler JD, Sokol J, Comly ME, Vanier MT, August JT, Brady RO & Pentchev PG 1988 Type C Niemann-Pick disease: Low density lipoprotein uptake is associated with premature cholesterol accumulation in the Golgi complex and excessive cholesterol storage in lysosomes. Proc Natl Acad Sci USA 85 8022-8026

Brown MS & Goldstein JL 1975 Regulation of the activity of the low density lipoprotein receptor in human fibroblasts. Cell 6 307-316

Brown MS, Ho YK & Goldstein JL 1976 The low-density lipoprotein pathway in human fibroblasts: Relation between cell surface receptor binding and endocytosis of low-density lipoprotein. Ann NY Acad Sci 275 244-257

Cheruku SR, Xu Z, Dutia R, Lobel P & Storch J 2006 Mechanism of cholesterol transfer from the Niemann-Pick type C2 protein to model membranes supports a role in lysosomal cholesterol transport. J Biol Chem 281 31594-31604

Couet J, Li S, Okamoto T, Ikezu T & Lisanti MP 1997 Identification of peptide and protein ligands for the caveolin-scaffolding domain. J Biol Chem 272 6525-6533

Coxey RA, Pentchev PG, Campbell G & Blanchette-Mackie EJ 1993 Differential accumulation of cholesterol in Golgi compartments of normal and Niemann-Pick type C fibroblasts incubated with LDL: A cytochemical freeze-fracture study. J Lipid Res 34 1165-1176

Dietschy JM, Turley SD, & Spady DK 1993 Role of liver in the maintenance of cholesterol and low density lipoprotein homeostasis in different animal species, including humans. J Lipid Res 34 1637-1659

Dupree P, Parton RG, Raposo G, Kurzchalia TV & Simons K 1993 Caveolae and sorting in the trans-Golgi network of epithelial cells. EMBO J 12 1597-1605

Fielding PE & Fielding CJ 1996 Intracellular transport of low density lipoprotein derived free cholesterol begins at clathrin-coated pits and terminates at cell surface caveolae. Biochemistry 35 14932-14938

Friedland N, Liou H-L, Lobel P & Stock AM 2003 Structure of a cholesterol-binding protein deficient in Niemann-Pick type C2 disease. Proc Nat Acad Sci USA 100 2512-2517

Garver WS, Erickson RP, Wilson JM, Colton TL, Hossain GS, Kozloski MA & Heidenreich RA 1997a Altered expression of caveolin-1 and increased cholesterol in detergent insoluble membrane fractions from liver in mice with Niemann-Pick disease type C. Biochim Biophys Acta 1361 272-280

Garver WS, Heidenreich RA, Erickson RP, Thomas MA & Wilson JM 2000 Localization of the murine Niemann-Pick C1 protein to two distinct intracellular compartments. J Lipid Res 41 673-687

Garver WS, Hsu SC, Erickson RP, Greer WL, Byers DM & Heidenreich RA 1997b Increased expression of caveolin-1 in heterozygous Niemann-Pick type II human fibroblasts. Biochem Biophys Res Comm 236 189-193

Garver WS, Jelinek D, Oyarzo JN, Flynn J, Zuckerman M, Krishnan K, Chung BH & Heidenreich RA 2007 Characterization of liver disease and lipid metabolism in the Niemann-Pick C1 mouse. J Cell Biochem 101 498-516

Garver WS, Krishnan K, Gallagos JR, Michikawa M, Francis GA & Heidenreich RA 2002 Niemann-Pick C1 protein regulates cholesterol transport to the trans-Golgi network and plasma membrane caveolae. J Lipid Res 43 579-589

Garver WS, Jelinek D, Francis GA & Murphy BD 2008 The Niemann-Pick C1 gene is downregulated by feedback inhibition of the SREBP pathway in humans fibroblasts. J Lipid Res 49 1090-1102

Goldstein JL, Dana SE, Faust JR, Beaudet AL & Brown MS 1975 Role of lysosomal acid lipase in the metabolism of plasma low density lipoprotein. Observations in cultured fibroblasts from a patient with cholesteryl ester storage disease. J Biol Chem 250 8487-8495

Hailstone D, Sleer LS, Parton RG & Stanley KK 1998 Regulation of caveolin and caveolae by cholesterol in MDCK cells. J Lipid Res 39 369-379

Higgins ME, Davies JP, Chen FW & Ioannou YA 1999 Niemann-Pick C1 is a late endosome-resident protein that transiently associates with lysosomes and the trans-Golgi network. Mol Gene Metab 68 1-13

Infante RE, Radhakrishnan A, Abi-Mosleh L, Kinch LN, Wang ML, Grishin NV, Goldstein JL & Brown MS 2008a Purified NPC1 Protein II. Localization of sterol binding to a 240-amino acid soluble luminal loop. J Biol Chem 283 1064-1075

Infante RE, Wang ML, Radhakrishnan A, Kwon HJ, Brown MS & Goldstein JL 2008b NPC2 facilitates bidirectional transfer of cholesterol between NPC1 and lipid bilayers, a step in cholesterol egress from lysosomes. Proc Natl Acad Sci USA 105 15287-15292

Karpen HE, Bukowski JT, Hughes T, Gratton J, Sessa WC & Gailani MR 2001 The sonic hedgehog receptor patched associates with caveolin-1 in cholesterol-rich microdomains of the plasma membrane. J Biol Chem 276 19503-19511

Kim CA, Delepin M, Boutet M, El Mourabit H, Le Lay S, Meier M, Nemani M, Bridel E, Leite CC, Bertola DR, Semple RK, O'Rahilly S, Dugail I, Capeau J, Lathrop M & Magre J 2008 Association of a homozygous nonsense caveolin mutation with Berardinelli-Seip congenital lipodystrophy. J Clin Endo Metab 93 1129-1134

Klein A, Amigo L, Retamal MJ, Morales MG, Miquel JF, Rigotti A & Zanlungo S 2006 NPC2 is expressed in human and murine liver and secreted into bile: Potential implications for body cholesterol homeostasis. Hepatology 43 126-133

Ko DC, Binkley J, Sidow A & Scott MP 2003 The integrity of a cholesterol-binding pocket in Niemann-Pick C2 protein is necessary to control lysosome cholesterol levels. Proc Natl Acad Sci USA 100 2518-2525

Kruth HS, Comly ME, Butler JD, Vanier MT, Fink JK, Wenger DA, Patel S & Pentchev PG 1986 Type C Niemann-Pick disease. Abnormal metabolism of low density lipoprotein in homozygous and heterozygous fibroblasts. J Biol Chem 261 16769-16774

Kwon HJ, Abi-Mosleh, Wang ML, Deisenhofer J, Goldstein JL, Brown MS & Infante RE 2009 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol Cell 137 1213-1224

Li H & Papadopoulos V 1998 Peripheral-type benzodiazepine receptor function in cholesterol transport. Identification of a putative cholesterol recognition/interaction amino acid sequence and consensus pattern. Endocrinology 139 4991-4997

Liscum L, Ruggiero RM & Faust JR 1989 The intracellular transport of low density lipoprotein-derived cholesterol is defective in Niemann-Pick type C fibroblasts. J Cell Biol 108 1625-1636

Liu R, Lu P, Chu JWK & Sharom FJ 2009 Characterization of fluorescent sterol binding to purified human NPC1. J Biol Chem 284 1840-1852

Murata M, Peranen J, Schreiner R, Wieland F, Kurzchalia TV & Simons K 1995 VIP21/caveolin is a cholesterol-binding protein. Proc Natl Acad Sci USA 92 10339-10343

Neufeld EB, Wastney M, Patel S, Suresh S, Conney AM, Dwyer NK, Roff CF, Ohno K, Morris JA, Carstea ED, Incardona JP, Strauss JF, Vanier MT, Patterson MC, Brady RO, Pentchev PG & Blanchette-Mackie EJ 1999 The Niemann-Pick C1 protein resides in a vesicular compartment linked to retrograde transport of multiple lysosomal cargo. J Biol Chem 274 9627-9635

Ohgami N, Ko DC, Thomas M, Scott MP, Chang CCY & Chang TY 2004 Binding between the Niemann-Pick C1 protein and a photoactivatable cholesterol analog requires a functional sterol-sensing domain. Proc Natl Acad Sci USA 101 12473-12478

Osono Y, Woollett LA, Herz J & Dietschy JM 1995 Role of the low density lipoprotein receptor in the flux of cholesterol through the plasma and across the tissues of the mouse. J Clin Invest 95 1124-1132

Pelkmans L, Burli T, Zerial M & Helenius A 2004 Caveolin-stabilized membrane domains as multifunctional transport and sorting devices in endocytic membrane traffic. Cell 118 767-780

Pentchev PG, Kruth HS, Comly ME, Butler JD, Vanier MT, Wenger DA & Patel S 1986 Type C Niemann-Pick disease. A parallel loss of regulatory responses in both the uptake and esterification of low density lipoprotein-derived cholesterol in cultured fibroblasts. J Biol Chem 261 16775-16780

Pol A, Luetterforst R, Lindsay M, Heino S, Ikonen E & Parton RG 2001 A caveolin dominant negative mutant associates with lipid bodies and induces intracellular cholesterol imbalance. J Cell Biol 152 1057-1070

Razani B, Engelman JA, Wang XB, Schubert W, Zhang XL, Marks CB, Macaluso F, Russell RG, Li M, Pestell RG, Di Vizio D, Hou H, Kneitz B, Lagaud G, Christ GJ, Edelmann W & Lisanti MP 2001 Caveolin-1 null mice are viable but show evidence of hyperproliferative and vascular abnormalities. J Biol Chem 276 38121-38138

Razani B, Combs TP, Wang XB, Frank PG, Park DS, Russell RG, Li M, Tang B, Jelicks LA, Scherer PE & Lisanti MP 2002 Caveolin-1-deficient mice are lean, resistant to diet-induced obesity, and show hypertriglyceridemia with adipocyte abnormalities. J Biol Chem 277 8635-8647

Roy S, Luetterforst R, Harding A, Apolloni A, Etheridge M, Stang E, Rolls B, Hancock JF & Parton RG 1999 Dominant-negative caveolin inhibits H-Ras function by disrupting cholesterol-rich plasma membrane domains. Nat Cell Biol 1 98-105

Sleat DE, Wiseman JA, El-Banna M, Price SM, Verot L, Shen MM, Tint GS, Vanier MT, Walkley SU & Lobel P 2004 Genetic evidence for nonredundant functional cooperativity between NPC1 and NPC2 in lipid transport. Proc Natl Acad Sci USA 101 5886-5891

Smart EJ, Ying Y, Donzell WC & Anderson RGW 1996 A role for caveolin in transport of cholesterol from endoplasmic reticulum to plasma membrane. J Biol Chem 271 29427-29435

Sugii S, Reid PC, Ohgami N, Du H & Chang TY 2003 Distinct endosomal compartments in early trafficking of low density lipoprotein-derived cholesterol. J Biol Chem 278 27180-27189

Turley SD, Spady DK & Dietschy JM 1995 Role of liver in the synthesis of cholesterol and the clearance of low density lipoproteins in the cynomolgus monkey. J Lipid Res 36 67-79

Uittenbogaard A, Everson WV, Matveev SV & Smart EJ 2002 Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-protein complex. J Biol Chem 277 4925-4931

Uittenbogaard A, Ying Y & Smart EJ 1998 Characterization of a cytosolic heat-shock protein-caveolin chaperone complex. Involvement in cholesterol trafficking. J Biol Chem 273 6525-6532

Underwood KW, Jacobs NL, Howley A & Liscum L 1998 Evidence for a cholesterol transport pathway from lysosomes to endoplasmic reticulum that is independent of the plasma membrane. J Biol Chem 273 4266-4274

Urano Y, Watanabe H, Murphy SR, Shibuya Y, Geng Y, Peden AA, Chang CCY & Chang TY 2008 Transport of LDL-derived cholesterol from the NPC1 compartment to the ER involves the trans-Golgi network and the SNARE protein complex. Proc Natl Acad Sci USA 105 16513-16518

Watari H, Blanchette-Mackie EJ, Dwyer NK, Glick JM, Patel S, Neufeld EB, Brady RO, Pentchev PG & Strauss III JF 1999a Niemann-Pick C1 protein: Obligatory roles for N-terminal domains and lysosomal targeting in cholesterol mobilization. Proc Natl Acad Sci USA 96 805-810

Watari H, Blanchette-Mackie EJ, Dwyer NK, Watari M, Neufeld EB, Patel S, Pentchev PG & Strauss III JF 1999b Mutations in the leucine zipper motif and sterol-sensing domain inactivate the Niemann-Pick C1 glycoprotein. J Biol Chem 274 21861-21866

Wojtanik KM & Liscum L 2003 The transport of low density lipoprotein-derived cholesterol to the plasma membrane is defective in NPC1 cells. J Biol Chem 278 14850-14856

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