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Journal of Lipid Research, Vol 36, 1211-1226, Copyright © 1995 by Lipid Research, Inc.
EJ Blanchette-Mackie, NK Dwyer, T Barber, RA Coxey, T Takeda, CM Rondinone, JL Theodorakis, AS Greenberg and C Londos
Immunocytochemistry was used to determine the intracellular location of
perilipins in adipocytes and the occurrence of these proteins in tissues
involved in triacylglycerol metabolism. Confocal microscopy and
3-dimensional analysis of 3T3-L1 adipocytes showed that perilipin
immunofluorescence, present on the surfaces of all sized lipid droplets,
appeared unevenly dispersed on the surfaces of many large lipid droplets.
Electron microscopy revealed that immunogold staining for perilipin was
located directly on the surface layer apposed to and surrounding the core
triacylglycerol of intracellular lipid droplets of adipocytes in culture or
from white and brown adipose tissue. Freeze- fracture electron microscopy
indicated that the hydrophobic face of this surface monolayer contained
particles identical in size and distribution to intramembranous particles
(IMPs), which are unique structural features of the hydrophobic faces of
bilayered membranes. Also, freeze-fracture replicas revealed areas of
continuity between the surface layer of lipid droplets and the membrane
leaflets of endoplasmic reticulum, suggesting that the droplet monolayer
surface is an area of endoplasmic reticulum membrane leaflet modified by
its unique content of perilipin. Microperoxisomes, identified by
immunostaining for catalase, were found closely associated with lipid
droplets, but external to and not in contact with the lipid droplet surface
layer. Vimentin, identified by immunofluorescence, was present around the
periphery of most lipid droplets in 3T3-L1 cells during early stages of
adipocyte development but, in contrast to perilipins, vimentin was not
around the periphery of many large lipid droplets in mature cells. Although
perilipin was at the surface of lipid droplets in adipocytes of lactating
mammary gland, none was found to be associated with the milk lipid droplets
in alveolar epithelial cells, nor was the protein found on the surfaces of
lipid droplets in hepatocytes. Studies in mammary gland show that perilipin
immunostaining will be a valuable tool for the identification of tissue
adipocytes severely depleted of their triacylglycerol stores and thus
without their characteristic spherical shape. Perilipin's singular location
on the surface monolayer of intracellular lipid droplets supports an
intimate role for the protein in the triacylglycerol metabolic functions of
adipocytes.
ARTICLES
Perilipin is located on the surface layer of intracellular lipid droplets in adipocytes
Lipid Cell Biology Section, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0850, USA.
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S. C. Souza, K. V. Muliro, L. Liscum, P. Lien, M. T. Yamamoto, J. E. Schaffer, G. E. Dallal, X. Wang, F. B. Kraemer, M. Obin, et al. Modulation of Hormone-sensitive Lipase and Protein Kinase A-mediated Lipolysis by Perilipin A in an Adenoviral Reconstituted System J. Biol. Chem., March 1, 2002; 277(10): 8267 - 8272. [Abstract] [Full Text] [PDF]
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N. B. Cole, D. D. Murphy, T. Grider, S. Rueter, D. Brasaemle, and R. L. Nussbaum Lipid Droplet Binding and Oligomerization Properties of the Parkinson's Disease Protein alpha -Synuclein J. Biol. Chem., February 15, 2002; 277(8): 6344 - 6352. [Abstract] [Full Text] [PDF]
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R. G. Hope, D. J. Murphy, and J. McLauchlan The Domains Required to Direct Core Proteins of Hepatitis C Virus and GB Virus-B to Lipid Droplets Share Common Features with Plant Oleosin Proteins J. Biol. Chem., February 1, 2002; 277(6): 4261 - 4270. [Abstract] [Full Text] [PDF]
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A. J. Charron and L. D. Sibley Host cells: mobilizable lipid resources for the intracellular parasite Toxoplasma gondii J. Cell Sci., January 8, 2002; 115(15): 3049 - 3059. [Abstract] [Full Text] [PDF]
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M. Schrader Tubulo-Reticular Clusters of Peroxisomes in Living COS-7 Cells: Dynamic Behavior and Association with Lipid Droplets J. Histochem. Cytochem., November 1, 2001; 49(11): 1421 - 1430. [Abstract] [Full Text] [PDF]
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A. Rudich, S. Vanounou, K. Riesenberg, M. Porat, A. Tirosh, I. Harman-Boehm, A. S. Greenberg, F. Schlaeffer, and N. Bashan The HIV Protease Inhibitor Nelfinavir Induces Insulin Resistance and Increases Basal Lipolysis in 3T3-L1 Adipocytes Diabetes, June 1, 2001; 50(6): 1425 - 1431. [Abstract] [Full Text]
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G. van Meer Caveolin, Cholesterol, and Lipid Droplets? J. Cell Biol., March 5, 2001; 152(5): F29 - F34. [Full Text] [PDF]
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T. Fujimoto, H. Kogo, K. Ishiguro, K. Tauchi, and R. Nomura Caveolin-2 Is Targeted to Lipid Droplets, a New "Membrane Domain" in the Cell J. Cell Biol., March 5, 2001; 152(5): 1079 - 1086. [Abstract] [Full Text] [PDF]
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G. M. Clifford, C. Londos, F. B. Kraemer, R. G. Vernon, and S. J. Yeaman Translocation of Hormone-sensitive Lipase and Perilipin upon Lipolytic Stimulation of Rat Adipocytes J. Biol. Chem., February 18, 2000; 275(7): 5011 - 5015. [Abstract] [Full Text] [PDF]
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S Prattes, G Horl, A Hammer, A Blaschitz, W. Graier, W Sattler, R Zechner, and E Steyrer Intracellular distribution and mobilization of unesterified cholesterol in adipocytes: triglyceride droplets are surrounded by cholesterol-rich ER-like surface layer structures J. Cell Sci., January 9, 2000; 113(17): 2977 - 2989. [Abstract] [PDF]
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R. Li, E. J. Blanchette-Mackie, and S. Ladisch Induction of Endocytic Vesicles by Exogenous C6-ceramide J. Biol. Chem., July 23, 1999; 274(30): 21121 - 21127. [Abstract] [Full Text] [PDF]
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L. S. Szczepaniak, E. E. Babcock, F. Schick, R. L. Dobbins, A. Garg, D. K. Burns, J. D. McGarry, and D. T. Stein Measurement of intracellular triglyceride stores by H spectroscopy: validation in vivo Am J Physiol Endocrinol Metab, May 1, 1999; 276(5): E977 - E989. [Abstract] [Full Text] [PDF]
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V. A. Barr, D. Malide, M. J. Zarnowski, S. I. Taylor, and S. W. Cushman Insulin Stimulates Both Leptin Secretion and Production by Rat White Adipose Tissue Endocrinology, October 1, 1997; 138(10): 4463 - 4472. [Abstract] [Full Text] [PDF]
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D. L. Brasaemle, T. Barber, A. R. Kimmel, and C. Londos Post-translational Regulation of Perilipin Expression. STABILIZATION BY STORED INTRACELLULAR NEUTRAL LIPIDS J. Biol. Chem., April 4, 1997; 272(14): 9378 - 9387. [Abstract] [Full Text] [PDF]
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D Malide and S. Cushman Morphological effects of wortmannin on the endosomal system and GLUT4-containing compartments in rat adipose cells J. Cell Sci., January 11, 1997; 110(22): 2795 - 2806. [Abstract] [PDF]
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J. Lieber and R. Evans Disruption of the vimentin intermediate filament system during adipose conversion of 3T3-L1 cells inhibits lipid droplet accumulation J. Cell Sci., January 12, 1996; 109(13): 3047 - 3058. [Abstract] [PDF]
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N. E. Wolins, B. Rubin, and D. L. Brasaemle TIP47 Associates with Lipid Droplets J. Biol. Chem., February 9, 2001; 276(7): 5101 - 5108. [Abstract] [Full Text] [PDF]
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D. L. Brasaemle, B. Rubin, I. A. Harten, J. Gruia-Gray, A. R. Kimmel, and C. Londos Perilipin A Increases Triacylglycerol Storage by Decreasing the Rate of Triacylglycerol Hydrolysis J. Biol. Chem., December 1, 2000; 275(49): 38486 - 38493. [Abstract] [Full Text] [PDF]
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S. Sonda, L.-M. Ting, S. Novak, K. Kim, J. J. Maher, R. V. Farese Jr., and J. D. Ernst Cholesterol Esterification by Host and Parasite Is Essential for Optimal Proliferation of Toxoplasma gondii J. Biol. Chem., September 7, 2001; 276(37): 34434 - 34440. [Abstract] [Full Text] [PDF]
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 B. C.G. Faber, K. B.J.M. Cleutjens, R. L.J. Niessen, P. L.J.W. Aarts, W. Boon, A. S. Greenberg, P. J.E.H.M. Kitslaar, J. H.M. Tordoir, and M. J.A.P. Daemen Identification of Genes Potentially Involved in Rupture of Human Atherosclerotic Plaques Circ. Res., September 14, 2001; 89(6): 547 - 554. [Abstract] [Full Text] [PDF]
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