Physical association between the adipocyte fatty acid-binding protein and hormone-sensitive lipase - A fluorescence resonance energy transfer analysis JOURNAL OF BIOLOGICAL CHEMISTRY SMITH, A. J., Sanders, M. A., Thompson, B. R., Londos, C., Kraemer, F. B., Bernlohr, D. A. 2004; 279 (50): 52399-52405


Previous in vitro studies have established that hormone sensitive lipase (HSL) and adipocyte fatty acid-binding protein (AFABP) form a physical complex that presumably positions the FABP to accept a product fatty acid generated during catalysis. To assess AFABP-HSL interaction within a cellular context, we have used lipocytes derived from 293 cells (C8PA cells) and examined physical association using fluorescence resonance energy transfer. Transfection of C8PA cells with cyan fluorescent protein (CFP)-HSL, yellow fluorescent protein (YFP)-adipocyte FABP, or YFP-liver FABP revealed that under basal conditions each protein was cytoplasmic. In the presence of 20 microm forskolin, CFP-HSL translocated to the triacylglycerol droplet, coincident with BODIPY-FA labeled depots. Fluorescence resonance energy transfer analysis demonstrated that CFP-HSL associated with YFP-adipocyte FABP in both basal and forskolin-treated cells. In contrast, little if any fluorescence resonance energy transfer could be detected between CFP-HSL and YFP-liver FABP. These results suggest that a pre-lipolysis complex containing at least AFABP and HSL exists and that the complex translocates to the surface of the lipid droplet.

View details for DOI 10.1074/jbc.M410301200

View details for Web of Science ID 000225493400084

View details for PubMedID 15456755