Document Detail

A phospholipase D-dependent process forms lipid droplets containing caveolin, adipocyte differentiation-related protein, and vimentin in a cell-free system.
MedLine Citation:
PMID:  12730229     Owner:  NLM     Status:  MEDLINE    
We developed a microsome-based, cell-free system that assembles newly formed triglyceride (TG) into spherical lipid droplets. These droplets were recovered in the d </= 1.055 g/ml fraction by gradient ultracentrifugation and were similar in size and appearance to those isolated from rat adipocytes and 3T3-L1 cells. Caveolin 1 and 2, vimentin, adipocyte differentiation-related protein, and the 78-kDa glucose regulatory protein were identified on the droplets from the cell-free system. The caveolin was soluble in 1% Triton X-100, as was the caveolin on lipid droplets from 3T3-L1 cells. The lipid droplets from the cell-free system, like those from 3T3-L1 cells, contained TG, diacylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine. The assembly of these TG-containing structures was dependent on the rate of TG biosynthesis and required an activator present in the 160,000 x g supernatant from homogenized rat adipocytes. The activator induced phospholipase D (PLD) activity, and its effect on the release of the TG-containing structures from the microsomes was inhibited by 1-butanol (but not 2-butanol) or 2,3-diphosphoglycerate. The activator could be replaced by a constitutively active PLD or phosphatidic acid. These results indicate that PLD and the formation of phosphatidic acid are important in the assembly of the TG-containing structures.
Denis Marchesan; Mikael Rutberg; Linda Andersson; Lennart Asp; Thomas Larsson; Jan Borén; Bengt R Johansson; Sven-Olof Olofsson
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't     Date:  2003-04-30
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  278     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-07-14     Completed Date:  2003-08-27     Revised Date:  2011-10-18    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  27293-300     Citation Subset:  IM    
Department of Medical Biochemistry and the Wallenberg Laboratory for Cardiovascular Research, Göteborg University, SE-413 45 Göteborg, Sweden.
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MeSH Terms
3T3 Cells
Adipocytes / metabolism
Caveolin 1
Caveolin 2
Caveolins / metabolism*
Cell-Free System
Lipid Metabolism*
Membrane Proteins / metabolism*
Microscopy, Electron
Microsomes / metabolism
Phospholipase D / metabolism*
Triglycerides / metabolism
Vimentin / metabolism*
Reg. No./Substance:
0/Cav protein, rat; 0/Cav1 protein, mouse; 0/Caveolin 1; 0/Caveolin 2; 0/Caveolins; 0/Membrane Proteins; 0/Triglycerides; 0/Vimentin; 0/perilipin 2; EC D

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