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Apolipoprotein A-V association with intracellular lipid droplets
Authors:Shu Xiao  Chan Joanne  Ryan Robert O  Forte Trudy M
Institution:Center for Prevention of Obesity, Diabetes, and Cardiovascular Diseases, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA.
Abstract:Apolipoprotein A-V (apoA-V) plays a key role in the regulation of triglyceride (TG) metabolism. Given the very low concentration of apoA-V in plasma, we hypothesized that apoA-V may influence plasma TG levels by affecting the assembly and/or secretion of apoB-containing lipoproteins. When apoA-V was overexpressed in cultured Hep3B cells, neither the amount of apoB secreted nor the density distribution of apoB-containing lipoproteins was affected. Fluorescence microscopy and cell lysate immunoprecipitation studies revealed that apoA-V is not associated with apoB intracellularly, yet immunoprecipitation of apoA-V from the cell culture medium resulted in coprecipitation of apoB. These data suggest that the apoA-V association with apoB-containing lipoproteins is a postsecretory event. Confocal fluorescence microscopy revealed the presence of apoA-V in distinct cellular structures. Based on Nile Red staining, we identified these structures to be intracellular lipid droplets. These data suggest that apoA-V has a unique association with cellular lipids and, therefore, may be involved in the storage or mobilization of intracellular lipids.
Keywords:Hep3B hepatoma cell line  triglyceride  very low density lipoprotein  immunoprecipitation  confocal fluorescence microscopy  apolipoprotein B
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