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Journal of Lipid Research, Vol. 48, 609-620, March 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Fatty acid metabolism in adipocytes: functional analysis of fatty acid transport proteins 1 and 4

Sandra Lobo, Brian M. Wiczer, Ann J. Smith, Angela M. Hall and David A. Bernlohr¹

Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455

Published, JLR Papers in Press, December 11, 2006.

¹ To whom correspondence should be addressed. e-mail: bernl001{at}umn.edu

The role of fatty acid transport protein 1 (FATP1) and FATP4 in facilitating adipocyte fatty acid metabolism was investigated using stable FATP1 or FATP4 knockdown (kd) 3T3-L1 cell lines derived from retrovirus-delivered short hairpin RNA (shRNA). Decreased expression of FATP1 or FATP4 did not affect preadipocyte differentiation or the expression of FATP1 (in FATP4 kd), FATP4 (in FATP1 kd), fatty acid translocase, acyl-coenzyme A synthetase 1, and adipocyte fatty acid binding protein but did lead to increased levels of peroxisome proliferator-activated receptor and CCAAT/enhancer binding protein . Both FATP1 and FATP4 kd adipocytes exhibited reduced triacylglycerol deposition and corresponding reductions in diacylglycerol and monoacylglycerol levels compared with control cells. FATP1 kd adipocytes displayed an 25% reduction in basal ³H-labeled fatty acid uptake and a complete loss of insulin-stimulated ³H-labeled fatty acid uptake compared with control adipocytes. In contrast, FATP4 kd adipocytes as well as HEK-293 cells overexpressing FATP4 did not display any changes in fatty acid influx. FATP4 kd cells exhibited increased basal lipolysis, whereas FATP1 kd cells exhibited no change in lipolytic capacity. Consistent with reduced triacylglycerol accumulation, FATP1 and FATP4 kd adipocytes exhibited enhanced 2-deoxyglucose uptake compared with control adipocytes. These findings define unique and distinct roles for FATP1 and FATP4 in adipose fatty acid metabolism.

Supplementary key words fatty acid influx • basal lipolysis • triacylglycerol synthesis • acyl-coenzyme A synthetase • 2-deoxyglucose uptake

Abbreviations: ACSL1, acyl-coenzyme A synthetase 1; AFABP/aP2, adipocyte fatty acid binding protein; CD36, fatty acid translocase; C/EBP, CCAAT/enhancer binding protein ; FATP1, fatty acid transport protein 1; kd, knockdown; KRH, Krebs-Ringer's HEPES; LCFA, long-chain fatty acid; PPAR, peroxisome proliferator-activated receptor ; RNAi, RNA interference; shRNA, short hairpin RNA

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