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Originally published as MBC in Press, 10.1091/mbc.E04-07-0616 on October 20, 2004

Vol. 16, Issue 1, 24-31, January 2005

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FAT/CD36-mediated Long-Chain Fatty Acid Uptake in Adipocytes Requires Plasma Membrane Rafts

Jürgen Pohl, Axel Ring, Ümine Korkmaz, Robert Ehehalt, and Wolfgang Stremmel *

Department of Internal Medicine IV, Ruprechts-Karls-University, D-69117 Heidelberg, Germany

Submitted July 22, 2004; Revised September 15, 2004; Accepted September 27, 2004
Monitoring Editor: Guido Guidotti

We previously reported that lipid rafts are involved in long-chain fatty acid (LCFA) uptake in 3T3-L1 adipocytes. The present data show that LCFA uptake does not depend on caveolae endocytosis because expression of a dominant negative mutant of dynamin had no effect on uptake of [3H]oleic acid, whereas it effectively prevented endocytosis of cholera toxin. Isolation of detergent-resistant membranes (DRMs) from 3T3-L1 cell homogenates revealed that FAT/CD36 was expressed in both DRMs and detergent-soluble membranes (DSMs), whereas FATP1 and FATP4 were present only in DSMs but not DRMs. Disruption of lipid rafts by cyclodextrin and specific inhibition of FAT/CD36 by sulfo-N-succinimidyl oleate (SSO) significantly decreased uptake of [3H]oleic acid, but simultaneous treatment had no additional or synergistic effects, suggesting that both treatments target the same mechanism. Indeed, subcellular fractionation demonstrated that plasma membrane fatty acid translocase (FAT/CD36) is exclusively located in lipid rafts, whereas intracellular FAT/CD36 cofractionated with DSMs. Binding assays confirmed that [3H]SSO predominantly binds to FAT/CD36 within plasma membrane DRMs. In conclusion, our data strongly suggest that FAT/CD36 mediates raft-dependent LCFA uptake. Plasma membrane lipid rafts might control LCFA uptake by regulating surface availability of FAT/CD36.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E04–07–0616. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E04-07-0616.

Abbreviations used: CD, cyclodextrin; DRM, detergent-resistant membrane; DSM, detergent-soluble membrane; ER, endoplasmic reticulum; LCFA, long-chain fatty acid; FABPpm, plasma membrane fatty acid binding protein; FATP, fatty acid transport protein; FAT/CD36, fatty acid translocase; SSO, sulfo-N-succinimidyl oleate.

* Corresponding author. E-mail address: wolfgang.stremmel{at}med.uniheidelberg.de.




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