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Title: Characterization of the proteome of cytoplasmic lipid droplets in mouse enterocytes after a dietary fat challenge

Author
item D'AGUILA, TERESA - Purdue University
item SIROHI, DEVIKA - Purdue University
item GRABOWSKI, JEFFREY - Purdue University
item HEDRICK, VICTORIA - Purdue University
item PAUL, LAKE - Purdue University
item GREENBERG, ANDREW - Jean Mayer Human Nutrition Research Center On Aging At Tufts University
item KUHN, RICHARD - Purdue University
item BUHMAN, KIMBERLY - Purdue University

Submitted to: PLOS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/8/2015
Publication Date: 5/18/2015
Citation: D'Aguila, T., Sirohi, D., Grabowski, J.M., Hedrick, V.E., Paul, L.N., Greenberg, A., Kuhn, R.J., Buhman, K.K. 2015. Characterization of the proteome of cytoplasmic lipid droplets in mouse enterocytes after a dietary fat challenge. PLoS One. 10(5):e0126823. https://doi.org/10.1371/journal.pone.0126823.

Interpretive Summary: When fat is ingested in our diet, some fat is rapidly absorbed through the intestine, and some fat is stored within intestinal cells within specific intracellular compartments called lipid droplets. The fat stored within lipid droplets is thought to be slowly broken down over hours and absorbed into the body. Regulation of storage and breakdown within lipid droplets of intestinal cells is thought to influence how efficiently fat is absorbed into the body. In this paper we identified about 37 proteins that specifically localize to lipid droplets. In future studies we will investigate the relative roles of these proteins to gain insight into how dietary fat is absorbed as well as the proteins' effect on modulating the rate at which fat is absorbed. The importance of this study is that the proteins identified will facilitate future studies to elucidate potential mechanisms by which fat absorption can be modulated to match the metabolic demands of the body.

Technical Abstract: Dietary fat absorption by the small intestine is a multistep process that regulates the uptake and delivery of essential nutrients and energy. One step of this process is the temporary storage of dietary fat in cytoplasmic lipid droplets (CLDs). The storage and mobilization of dietary fat is thought to be regulated by proteins that associate with the CLD; however, mechanistic details of this process are currently unknown. In this study we analyzed the proteome of CLDs isolated from enterocytes harvested from the small intestine of mice following a dietary fat challenge. In this analysis we identified 181 proteins associated with the CLD fraction, of which 37 are associated with known lipid related metabolic pathways. We confirmed the localization of several of these proteins on or around the CLD through confocal and electron microscopy, including perilipin 3, apolipoprotein A-IV, and acyl-CoA synthetase long-chain family member 5. The identification of the enterocyte CLD proteome provides new insight into potential regulators of CLD metabolism and the process of dietary fat absorption.