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Papers In Press, published online ahead of print July 1, 2007
J. Lipid Res., doi:10.1194/jlr.C700002-JLR200

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Journal of Lipid Research, Vol. 48, 1445-1450, July 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Short Communication

Apolipoprotein A-V association with intracellular lipid droplets

Xiao Shu^*,, Joanne Chan^*, Robert O. Ryan^*, and Trudy M. Forte^1,*

^* Center for Prevention of Obesity, Diabetes, and Cardiovascular Diseases, Children's Hospital Oakland Research Institute, Oakland, CA 94609
Department of Nutritional Sciences and Toxicology, University of California, Berkeley, CA 94720

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of two figures.

Published, JLR Papers in Press, April 25, 2007.

¹ To whom correspondence should be addressed. e-mail: tforte{at}chori.org

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.

Supplementary key words Hep3B hepatoma cell line • triglyceride • very low density lipoprotein • immunoprecipitation • confocal fluorescence microscopy • apolipoprotein B

Abbreviations: apoA-V, apolipoprotein A-V; GFP, green fluorescent protein; Met/Cys, methionine/cysteine; OA, oleic acid; TG, triglyceride

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