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J. Biol. Chem., Vol. 275, Issue 49, 38486-38493, December 8, 2000
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From the The perilipins are the most abundant proteins at
the surfaces of lipid droplets in adipocytes and are also found in
steroidogenic cells. To investigate perilipin function, perilipin A,
the predominant isoform, was ectopically expressed in fibroblastic
3T3-L1 pre-adipocytes that normally lack the perilipins. In control
cells, fluorescent staining of neutral lipids with Bodipy 493/503
showed a few minute and widely dispersed lipid droplets, while in cells
stably expressing perilipin A, the lipid droplets were more numerous
and tightly clustered in one or two regions of the cytoplasm.
Immunofluorescence microscopy revealed that the ectopic perilipin A
localized to the surfaces of the tiny clustered lipid droplets;
subcellular fractionation of the cells using sucrose gradients
confirmed that the perilipin A localized exclusively to lipid droplets.
Cells expressing perilipin A stored 6-30-fold more triacylglycerol
than control cells due to reduced lipolysis of triacylglycerol stores. The lipolysis of stored triacylglycerol was 5 times slower in lipid-loaded cells expressing perilipin A than in lipid-loaded control
cells, when triacylglycerol synthesis was blocked with 6 µM triacsin C. This stabilization of
triacylglycerol was not due to the suppression of triacylglycerol
lipase activity by the expression of perilipin A. We conclude that
perilipin A increases the triacylglycerol content of cells by forming a
barrier that reduces the access of soluble lipases to stored
lipids, thus inhibiting triacylglycerol hydrolysis. These studies
suggest that perilipin A plays a major role in the regulation of
triacylglycerol storage and lipolysis in adipocytes.
Perilipin A Increases Triacylglycerol Storage by Decreasing the
Rate of Triacylglycerol Hydrolysis*
§¶,§,
**,, and
Department of Biochemistry, MCP Hahnemann
University, Philadelphia, Pennsylvania 19129, the
§ Department of Nutritional Sciences, Rutgers, The State
University of New Jersey, New Brunswick, New Jersey 08901, and the
Molecular Mechanisms of Development Section and
Membrane Regulation Section, Laboratory of
Cellular and Developmental Biology, NIDDK, National Institutes of
Health, Bethesda, Maryland 20892
*
This work was supported by American Heart
Association-Southeastern Pennsylvania Affiliate Grant B98429E (to
D. L. B.) and an American Diabetes Association research grant
(to D. L. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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D. Marchesan, M. Rutberg, L. Andersson, L. Asp, T. Larsson, J. Boren, B. R. Johansson, and S.-O. Olofsson A Phospholipase D-dependent Process Forms Lipid Droplets Containing Caveolin, Adipocyte Differentiation-related Protein, and Vimentin in a Cell-free System J. Biol. Chem., July 11, 2003; 278(29): 27293 - 27300. [Abstract] [Full Text] [PDF]
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 C. Sztalryd, G. Xu, H. Dorward, J. T. Tansey, J. A. Contreras, A. R. Kimmel, and C. Londos Perilipin A is essential for the translocation of hormone-sensitive lipase during lipolytic activation J. Cell Biol., June 23, 2003; 161(6): 1093 - 1103. [Abstract] [Full Text] [PDF]
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 D. DiDonato and D. L. Brasaemle Fixation Methods for the Study of Lipid Droplets by Immunofluorescence Microscopy J. Histochem. Cytochem., June 1, 2003; 51(6): 773 - 780. [Abstract] [Full Text] [PDF]
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P. Targett-Adams, D. Chambers, S. Gledhill, R. G. Hope, J. F. Coy, A. Girod, and J. McLauchlan Live Cell Analysis and Targeting of the Lipid Droplet-binding Adipocyte Differentiation-related Protein J. Biol. Chem., April 25, 2003; 278(18): 15998 - 16007. [Abstract] [Full Text] [PDF]
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J. T. Tansey, A. M. Huml, R. Vogt, K. E. Davis, J. M. Jones, K. A. Fraser, D. L. Brasaemle, A. R. Kimmel, and C. Londos Functional Studies on Native and Mutated Forms of Perilipins. A ROLE IN PROTEIN KINASE A-MEDIATED LIPOLYSIS OF TRIACYLGLYCEROLS IN CHINESE HAMSTER OVARY CELLS J. Biol. Chem., February 28, 2003; 278(10): 8401 - 8406. [Abstract] [Full Text] [PDF]
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A. Garcia, A. Sekowski, V. Subramanian, and D. L. Brasaemle The Central Domain Is Required to Target and Anchor Perilipin A to Lipid Droplets J. Biol. Chem., January 3, 2003; 278(1): 625 - 635. [Abstract] [Full Text] [PDF]
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H. H. Zhang, M. Halbleib, F. Ahmad, V. C. Manganiello, and A. S. Greenberg Tumor Necrosis Factor-{alpha} Stimulates Lipolysis in Differentiated Human Adipocytes Through Activation of Extracellular Signal-Related Kinase and Elevation of Intracellular cAMP Diabetes, October 1, 2002; 51(10): 2929 - 2935. [Abstract] [Full Text] [PDF]
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M. Imamura, T. Inoguchi, S. Ikuyama, S. Taniguchi, K. Kobayashi, N. Nakashima, and H. Nawata ADRP stimulates lipid accumulation and lipid droplet formation in murine fibroblasts Am J Physiol Endocrinol Metab, October 1, 2002; 283(4): E775 - E783. [Abstract] [Full Text] [PDF]
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S. Miura, J.-W. Gan, J. Brzostowski, M. J. Parisi, C. J. Schultz, C. Londos, B. Oliver, and A. R. Kimmel Functional Conservation for Lipid Storage Droplet Association among Perilipin, ADRP, and TIP47 (PAT)-related Proteins in Mammals, Drosophila, and Dictyostelium J. Biol. Chem., August 23, 2002; 277(35): 32253 - 32257. [Abstract] [Full Text] [PDF]
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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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T. Raclot, C. Holm, and D. Langin Fatty acid specificity of hormone-sensitive lipase: implication in the selective hydrolysis of triacylglycerols J. Lipid Res., December 1, 2001; 42(12): 2049 - 2057. [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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J. T. Tansey, C. Sztalryd, J. Gruia-Gray, D. L. Roush, J. V. Zee, O. Gavrilova, M. L. Reitman, C.-X. Deng, C. Li, A. R. Kimmel, et al. Perilipin ablation results in a lean mouse with aberrant adipocyte lipolysis, enhanced leptin production, and resistance to diet-induced obesity PNAS, May 22, 2001; 98(11): 6494 - 6499. [Abstract] [Full Text] [PDF]
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A. G. Ostermeyer, J. M. Paci, Y. Zeng, D. M. Lublin, S. Munro, and D. A. Brown Accumulation of Caveolin in the Endoplasmic Reticulum Redirects the Protein to Lipid Storage Droplets J. Cell Biol., March 5, 2001; 152(5): 1071 - 1078. [Abstract] [Full Text] [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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L. Izem and R. E. Morton Cholesteryl Ester Transfer Protein Biosynthesis and Cellular Cholesterol Homeostasis Are Tightly Interconnected J. Biol. Chem., July 6, 2001; 276(28): 26534 - 26541. [Abstract] [Full Text] [PDF]
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