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MBC in Press, published online ahead of print October 20, 2004
Mol. Biol. Cell 10.1091/mbc.E04-07-0616

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Submitted on July 22, 2004
Revised on September 15, 2004
Accepted on September 27, 2004

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

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 as expression of a dominant negative mutant of dynamin had no effect on uptake of [3H] oleic acid while 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) while FATP1 and FATP4 were present only in DSM 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 while 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.

*Corresponding author. E-mail: wolfgang.stremmel{at}med.uni-heidelberg.de




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