Expression of the CD36 homolog (FAT) in fibroblast cells: effects on fatty acid transport.
An adipocyte membrane glycoprotein, (FAT), homologous to human CD36, has been previously implicated in the binding/transport of long-chain fatty acids. It bound reactive derivatives of long-chain fatty acids and binding was specific and associated with significant inhibition of fatty acid uptake. Tissue distribution of the protein and regulation of its expression were also consistent with its postulated role. In this report, we have examined the effects of FAT expression on rates and properties of fatty acid uptake by Ob17PY fibroblasts lacking the protein. Three clones (P21, P22, and P25) were selected based on FAT mRNA and protein levels. Cell surface labeling could be demonstrated with the anti-CD36 antibody FITC-OKM5. In line with this, the major fraction of immunoreactive FAT was associated with the plasma membrane fraction. Assays of oleate and/or palmitate uptake demonstrated higher rates in the three FAT-expressing clones, compared to cells transfected with the empty vector. Clone P21, which had the highest protein levels on Western blots, exhibited the largest increase in transport rates. Fatty acid uptake in FAT-expressing P21 cells reflected two components, a phloretin-sensitive high-affinity saturable component with a Km of 0.004 microM and a basal phloretin-insensitive component that was a linear function of unbound fatty acid. P21 cells incorporated more exogenous fatty acid into phospholipids, indicating that binding of fatty acids was followed by their transfer into the cell and that both processes were increased by FAT expression. The data support the interpretation that FAT/CD36 functions as a high-affinity membrane receptor/transporter for long-chain fatty acids.
Ibrahimi
A
A
Department of Physiology and Biophysics, State University of New York at Stony Brook, 11733, USA.
Sfeir
Z
Z
Magharaie
H
H
Amri
E Z
EZ
Grimaldi
P
P
Abumrad
N A
NA
D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001692:Biological Transport; D018955:CD36 Antigens / Q000378:metabolism*; D002460:Cell Line; D000090266:Chromobox Protein Homolog 5; D002999:Clone Cells; D005227:Fatty Acids / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D027361:Organic Anion Transporters; D010693:Phloretin / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
D015703:Antigens, CD; C000715594:CBX5 protein, human; D018955:CD36 Antigens; D005227:Fatty Acids; D008562:Membrane Glycoproteins; D009829:Oleic Acids; D027361:Organic Anion Transporters; D011994:Recombinant Proteins; D000090266:Chromobox Protein Homolog 5; D019301:Oleic Acid; D010693:Phloretin