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Journal of Lipid Research, Vol. 43, 1380-1389, September 2002
Copyright © 2002 by Lipid Research, Inc.
* Departments of Pharmacology, University of Manitoba, Winnipeg, Manitoba, Canada
Therapeutics, University of Manitoba, Winnipeg, Manitoba, Canada
Department of Biochemistry, Molecular Biology and Biophysics, The University of Minnesota, Minneapolis, MN
1 To whom correspondence should be addressed. e-mail: bernl001{at}umn.edu
Biosynthesis of lipids was investigated in growing 293 cells stably expressing fatty acid (FA) transport protein 1 (FATP1), a bifunctional polypeptide with FA transport as well as fatty acyl-CoA synthetase activity. In short-term (30 s) incubations, FA uptake was increased in FATP1 expressing cells (C8 cells) compared with the vector (as determined by BODIPY 3823 staining and radioactive FA uptake). In long-term (4 h) incubations, incorporation of [14C]acetate, [3H]oleic acid, or [14C]lignoceric acid into 1,2,3-triacyl-sn-glycerol (TG) was elevated in C8 cells compared with vector, whereas incorporation of radiolabel into glycerophospholipids was unaltered. The increase in TG biosynthesis correlated with an increase in 1,2-diacyl-sn-glycerol acyltransferase activity in C8 cells compared with vector. In contrast, incorporation of [14C]acetate into sphingomyelin (SM) and cholesterol, and [3H]oleic acid or [14C]lignoceric acid into SM was reduced due to a reduction in de novo biosynthesis of these lipids in C8 cells compared with vector.
The results indicate that exogenously supplied FAs, and their subsequently produced acyl-CoAs, are preferentially channeled by an FATP1 linked mechanism into the TG biosynthetic pathway and that such internalized lipids down-regulate de novo SM and cholesterol metabolism in actively growing 293 cells.
Abbreviations: ACS, acyl-CoA synthetase; DG, 1,2-diacyl-sn-glycerol; DGAT, 1,2-diacyl-sn-glycerol acyltransferase; ER, endoplasmic reticulum; FA, fatty acid; FATP1, fatty acid transport protein 1; GPAT, sn-glycerol-3-phosphate acyltransferase; MHC, myosin heavy chain; PA, phosphatidic acid; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PM, plasma membrane; PS, phosphatidylserine; SM, sphingomyelin; TG, 1,2,3-triacyl-sn-glycerol
Supplementary key words transport • CoA synthetase • esterification • fatty acid transport protein 1
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