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Expression of rat L-FABP in mouse fibroblasts: role in fat absorption

  • Cellular Fatty Acid-Binding Proteins II
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Abstract

Fatty acid-binding proteins (FABP) are abundant cytosolic proteins whose level is responsive to nutritional, endocrine, and a variety of pathological states. Although FABPs have been investigatedin vitro for several decades, little is known of their physiological function. Liver L-FABP binds both fatty acids and cholesterol. Competitive binding analysis and molecular modeling studies of L-FABP indicate the presence of two ligand binding pockets that accomodate one fatty acid each. One fatty acid binding site is identical to the cholesterol binding site. To test whether these observations obtainedin vitro were physiologically relevant, the cDNA encoding L-FABP was transfected into L-cells, a cell line with very low endogenous FABP and sterol carrier proteins. Uptake of both ligands did not differ between control cells and low expression clones. In contrast, both fatty acid uptake and cholesterol uptake were stimulated in the high expression cells. In high expression cells, uptake of fluorescent cis-parinaric acid was enhanced more than that of trans-parinaric acid. This is consistent with the preferential binding of cis-fatty acids to L-FABP but in contrast to the preferential binding of trans-parinaric acid to the L-cell plasma membrane fatty acid transporter (PMFABP). These data show that the level of cytosolic fatty acids in intact cells can regulate both the extent and specificity of fatty acid uptake. Last, sphingomyelinase treatment of L-cells released cholesterol from the plasma membrane to the cytoplasm and stimulated microsomal acyl-CoA: cholesteryl acyl transferase (ACAT). This process was accelerated in high expression cells. These observations show for the first time in intact cells that L-FABP, a protein most prevalent in liver and intestine where much fat absorption takes place, may have a role in fatty acid and cholesterol absorption.

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Abbreviations

FABP:

fatty acid-binding protein

L-FABP:

liver fatty acid-binding protein

I-FABP:

intestinal fatty acid-binding protein

H-FABP:

heart fatty acid-binding protein

A-FABP:

adipocyte fatty acid-binding protein

PMFABP:

plasma membrane fatty acid-binding protein

SCP-2:

sterol carrier protein-2

Dehydroergosterol (DHE):

d-5,7,9(11),22-ergostatetraene-3b-ol

cis-parinaric:

acid-9Z, 11E, 13E, 15Z-octatetraenoic acid

trans:

parinaric acid, 9E, 11E, 13E, 14E-octatetraenoic acid

BSA:

bovine serum albumin

KRH:

Krebs-Ringer-Henseleit buffer

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Authors and Affiliations

  1. Divi of Pharmacology & Med. Chem., Dept. of Pharmacology & Cell Biophysics, University of Cincinnati Medical Center, ML004, 45267-0004, Cincinnati, OH, USA

    F. Schroeder, D. Powell, J. K. Woodford, S. M. Colles, S. Myers-Payne, T. Hubbell, D. Moncecchi, D. R. Prows & C. E. Heyliger

  2. Dept. of Chemistry, University of Cincinnati, 45267-0172, Cincinnati, OH, USA

    T. Emge

  3. Dept. of Chemistry, Luther College, 52101-1045, Decorah, IA, USA

    J. R. Jefferson

  4. Dipartimento Di Biologia, Universita Degli Studi Di Roma, Tor Vergata, Rome, Italy

    S. Incerpi

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  1. F. Schroeder
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  2. J. R. Jefferson
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  8. T. Emge
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Schroeder, F., Jefferson, J.R., Powell, D. et al. Expression of rat L-FABP in mouse fibroblasts: role in fat absorption. Mol Cell Biochem 123, 73–83 (1993). https://doi.org/10.1007/BF01076477

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