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Journal of Lipid Research, Vol. 44, 668-673, April 2003
Copyright © 2003 by Lipid Research, Inc.
Rapid Communication |
Lipid droplet targeting domains of adipophilin
* Department of Obstetrics and Gynecology, University of Colorado Health Sciences Center, Denver, CO 80261
Division of Basic Reproductive Science, Department of Physiology and Biophysics, University of Colorado Health Sciences Center, Denver, CO 80261
Department of Virology and Microbiology, University of Colorado Health Sciences Center, Denver, CO 80261
** Department of Pathology, University of Colorado Health Sciences Center, Denver, CO 80261
1 To whom correspondence should be addressed. e-mail: jim.mcmanaman{at}uchsc.edu
ABSTRACT
Adipophilin (ADPH), a prominent protein component of lipid storage droplets (LSDs), is postulated to be necessary for the formation and cellular function of these structures. The presence of significant sequence similarities within an 
100 amino acid region of the N-terminal portions of ADPH and related LSD binding proteins, perilipin and TIP47, has implicated this region, known as the "PAT" domain, in LSD targeting. Here we investigate the role of the PAT domain in targeting ADPH to LSDs by expressing this region, as well as selected N- and C-terminal truncations of mouse ADPH in COS7 cells as epitope-tagged fusion proteins. Our studies show that truncations lacking either the PAT domain or the C-terminal half of ADPH both correctly targeted LSDs and increased the LSD content of transfected cells. Neither the PAT domain nor the C-terminal half of ADPH appeared to target LSDs or affect the LSD number.
Instead, targeting fragments encompassed a putative 
-helical region between amino acids 189 and 205, implicating this region in both LSD targeting and regulation of LSD formation.
Abbreviations: ADPH, adipophilin; GRAVY, grand average hydrophobicity; LSD, lipid storage droplet; PAT, perilipin, adipophilin, TIP47
Supplementary key words lipid storage droplets • protein targeting • functional domains • cell culture • transfection • immunofluorescence
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