Document Detail


In vitro formation of a novel type of membrane vesicles containing Dpm1p: putative transport vesicles for lipid droplets in budding yeast.
MedLine Citation:
PMID:  18343875     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel type of membrane vesicles was formed in vitro from microsomes of Saccharomyces cerevisiae, which carries Dpm1p, an enzyme involved in dolichol-sugar synthesis, but not a typical secretory cargo. While COPII vesicles formed in vitro were sedimentable by centrifugation at 200,000g(max) for 15 min, the novel vesicles were not. However, they were sedimented by additional centrifugation at the same speed for 1 h. Immunoelectron microscopy showed that the Dpm1p-containing vesicles had small vesicular/saccular structures of around 40-50 nm in diameter. The addition of glycerol-3-phosphate and oleoyl-CoA, substrates for lipid biosynthesis, significantly enhanced the efficiency of vesicle budding in an ATP-dependent fashion. Dpm1p was localized to lipid droplets as well as endoplasmic reticulum. Fluorescence microscopy further showed that Dpm1p-GFP was present in restricted subregions in isolated lipid droplets. The possibility that the vesicles were intermediates from the endoplasmic reticulum to lipid droplets was examined, and their possible role is discussed.
Authors:
Yuichi Takeda; Akihiko Nakano
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Publication Detail:
Type:  Journal Article     Date:  2008-03-13
Journal Detail:
Title:  Journal of biochemistry     Volume:  143     ISSN:  0021-924X     ISO Abbreviation:  J. Biochem.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-06-27     Completed Date:  2008-09-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376600     Medline TA:  J Biochem     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  803-11     Citation Subset:  IM    
Affiliation:
Molecular Membrane Biology Laboratory, RIKEN Discovery Research Institute, 2-1 Hirosawa, Wako, Saitama 351-0198 Japan. y-takeda@biochem.med.osaka-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
COP-Coated Vesicles / metabolism*
Cytoplasm / metabolism
Golgi Apparatus / metabolism
Green Fluorescent Proteins / metabolism
Lipids / analysis*
Mannosyltransferases / genetics,  metabolism*
Microscopy, Fluorescence
Microsomes / metabolism*,  pathology
Saccharomycetales / metabolism*
Spheroplasts / metabolism
Subcellular Fractions
Chemical
Reg. No./Substance:
0/Lipids; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; EC 2.4.1.-/Mannosyltransferases; EC 2.4.1.83/dolichyl-phosphate beta-D-mannosyltransferase

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