Perilipin promotes hormone-sensitive lipase-mediated adipocyte lipolysis via phosphorylation-dependent and -independent mechanisms

Hideaki Miyoshi, Sandra C. Souza, Hui Hong Zhang, Katherine J. Strissel, Marcelo A. Christoffolete, Julia Kovsan, Assaf Rudich, Fredric B. Kraemer, Antonio C. Bianco, Martin S. Obin, Andrew S. Greenberg

Research output: Contribution to journalArticle

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Abstract

Hormone-sensitive lipase (HSL) is the predominant lipase effector of catecholamine-stimulated lipolysis in adipocytes. HSL-dependent lipolysis in response to catecholamines is mediated by protein kinase A (PKA)-dependent phosphorylation of perilipin A (Peri A), an essential lipid droplet (LD)-associated protein. It is believed that perilipin phosphorylation is essential for the translocation of HSL from the cytosol to the LD, a key event in stimulated lipolysis. Using adipocytes retrovirally engineered from murine embryonic fibroblasts of perilipin null mice (Peri-/- MEF), we demonstrate by cell fractionation and confocal microscopy that up to 50% of cellular HSL is LD-associated in the basal state and that PKA-stimulated HSL translocation is fully supported by adenoviral expression of a mutant perilipin lacking all six PKA sites (Peri AΔ1-6). PKA-stimulated HSL translocation was confirmed in differentiated brown adipocytes from perilipin null mice expressing an adipose-specific Peri AΔ1-6 transgene. Thus, PKA-induced HSL translocation was independent of perilipin phosphorylation. However, Peri AΔ1-6 failed to enhance PKA-stimulated lipolysis in either MEF adipocytes or differentiated brown adipocytes. Thus, the lipolytic action(s) of HSL at the LD surface requires PKA-dependent perilipin phosphorylation. In Peri-/- MEF adipocytes, PKA activation significantly enhanced the amount of HSL that could be cross-linked to and co-immunoprecipitated with ectopic Peri A. Notably, this enhanced cross-linking was blunted in Peri-/- MEF adipocytes expressing Peri AΔ1-6. This suggests that PKA-dependent perilipin phosphorylation facilitates (either direct or indirect) perilipin interaction with LD-associated HSL. These results redefine and expand our understanding of how perilipin regulates HSL-mediated lipolysis in adipocytes.

Original languageEnglish
Pages (from-to)15837-15844
Number of pages8
JournalJournal of Biological Chemistry
Volume281
Issue number23
DOIs
StatePublished - Jun 9 2006

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Sterol Esterase
Phosphorylation
Lipolysis
Adipocytes
Cyclic AMP-Dependent Protein Kinases
Lipids
Brown Adipocytes
Catecholamines
Perilipin-1
Cell Fractionation
Confocal microscopy
Fibroblasts
Fractionation
Lipase
Transgenes
Confocal Microscopy
Cytosol
Membrane Proteins
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Miyoshi, H., Souza, S. C., Zhang, H. H., Strissel, K. J., Christoffolete, M. A., Kovsan, J., ... Greenberg, A. S. (2006). Perilipin promotes hormone-sensitive lipase-mediated adipocyte lipolysis via phosphorylation-dependent and -independent mechanisms. Journal of Biological Chemistry, 281(23), 15837-15844. https://doi.org/10.1074/jbc.M601097200

Perilipin promotes hormone-sensitive lipase-mediated adipocyte lipolysis via phosphorylation-dependent and -independent mechanisms. / Miyoshi, Hideaki; Souza, Sandra C.; Zhang, Hui Hong; Strissel, Katherine J.; Christoffolete, Marcelo A.; Kovsan, Julia; Rudich, Assaf; Kraemer, Fredric B.; Bianco, Antonio C.; Obin, Martin S.; Greenberg, Andrew S.

In: Journal of Biological Chemistry, Vol. 281, No. 23, 09.06.2006, p. 15837-15844.

Research output: Contribution to journalArticle

Miyoshi, H, Souza, SC, Zhang, HH, Strissel, KJ, Christoffolete, MA, Kovsan, J, Rudich, A, Kraemer, FB, Bianco, AC, Obin, MS & Greenberg, AS 2006, 'Perilipin promotes hormone-sensitive lipase-mediated adipocyte lipolysis via phosphorylation-dependent and -independent mechanisms', Journal of Biological Chemistry, vol. 281, no. 23, pp. 15837-15844. https://doi.org/10.1074/jbc.M601097200
Miyoshi, Hideaki ; Souza, Sandra C. ; Zhang, Hui Hong ; Strissel, Katherine J. ; Christoffolete, Marcelo A. ; Kovsan, Julia ; Rudich, Assaf ; Kraemer, Fredric B. ; Bianco, Antonio C. ; Obin, Martin S. ; Greenberg, Andrew S. / Perilipin promotes hormone-sensitive lipase-mediated adipocyte lipolysis via phosphorylation-dependent and -independent mechanisms. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 23. pp. 15837-15844.
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abstract = "Hormone-sensitive lipase (HSL) is the predominant lipase effector of catecholamine-stimulated lipolysis in adipocytes. HSL-dependent lipolysis in response to catecholamines is mediated by protein kinase A (PKA)-dependent phosphorylation of perilipin A (Peri A), an essential lipid droplet (LD)-associated protein. It is believed that perilipin phosphorylation is essential for the translocation of HSL from the cytosol to the LD, a key event in stimulated lipolysis. Using adipocytes retrovirally engineered from murine embryonic fibroblasts of perilipin null mice (Peri-/- MEF), we demonstrate by cell fractionation and confocal microscopy that up to 50{\%} of cellular HSL is LD-associated in the basal state and that PKA-stimulated HSL translocation is fully supported by adenoviral expression of a mutant perilipin lacking all six PKA sites (Peri AΔ1-6). PKA-stimulated HSL translocation was confirmed in differentiated brown adipocytes from perilipin null mice expressing an adipose-specific Peri AΔ1-6 transgene. Thus, PKA-induced HSL translocation was independent of perilipin phosphorylation. However, Peri AΔ1-6 failed to enhance PKA-stimulated lipolysis in either MEF adipocytes or differentiated brown adipocytes. Thus, the lipolytic action(s) of HSL at the LD surface requires PKA-dependent perilipin phosphorylation. In Peri-/- MEF adipocytes, PKA activation significantly enhanced the amount of HSL that could be cross-linked to and co-immunoprecipitated with ectopic Peri A. Notably, this enhanced cross-linking was blunted in Peri-/- MEF adipocytes expressing Peri AΔ1-6. This suggests that PKA-dependent perilipin phosphorylation facilitates (either direct or indirect) perilipin interaction with LD-associated HSL. These results redefine and expand our understanding of how perilipin regulates HSL-mediated lipolysis in adipocytes.",
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AU - Christoffolete, Marcelo A.

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AU - Rudich, Assaf

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