Interaction of fatty acids with recombinant rat intestinal and liver fatty acid-binding proteins

Intestinal enterocytes contain two homologous fatty acid-binding proteins, intestinal fatty acid-binding protein (I-FABP)2 and liver fatty acid-binding protein (L-FABP). Since the functional basis for this multiplicity is not known, the fatty acid-binding specificity of recombinant forms of both rat I-FABP and rat L-FABP was examined. A systematic comparative analysis of the 18 carbon chain length fatty acid binding parameters, using both radiolabeled (stearic, oleic, and linoleic) and fluorescent (trans-parinaric and cis-parinaric) fatty acids, was undertaken. Results obtained with a classical Lipidex-1000 binding assay, which requires separation of bound from free fatty acid, were confirmed with a fluorescent fatty acid-binding assay not requiring separation of bound and unbound ligand. Depending on the nature of the fatty acid ligand, I-FABP bound fatty acid had dissociation constants between 0.2 and 3.1 [mu] and a consistent 1:1 molar ratio. The dissociation constants for L-FABP bound fatty acids ranged between 0.9 and 2.6 [mu] and the protein bound up to 2 mol fatty acid per mole of protein. Both fatty acid-binding proteins exhibited relatively higher affinity for unsaturated fatty acids as compared to saturated fatty acids of the same chain length. cis-Parinaric acid or trans-parinaric acid (each containing four double bonds) bound to L-FABP and I-FABP were displaced in a competitive manner by nonfluorescent fatty acid. Hill plots of the binding of cis- and trans- parinaric acid to L-FABP showed that the binding affinities of the two sites were very similar and did not exhibit cooperativity. The lack of fluorescence self-quenching upon binding 2 mol of either trans- or cis- parinaric acid/mol L-FABP is consistent with the presence of two binding sites with dissimilar orientation in the L-FABP. Thus, the difference in binding capacity between I-FABP and L-FABP predicts a structurally different binding site or sites.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/29398/1/0000471.pd

Bibliographie critique

Bastid Suzanne, Salmon Jean, Maisl Herbert, Fischer Georges, Pick H. H., Pellet Alain, Saint-Girons M.C., Depitre H., Sacchetini H., G. G., P. J.F., R. P. Bibliographie critique. In: Annuaire français de droit international, volume 14, 1968. pp. 971-1013

Bibliographie critique

Bastid Suzanne, Salmon Jean, Maisl Herbert, Fischer Georges, Pick H. H., Pellet Alain, Saint-Girons M.C., Depitre H., Sacchetini H., G. G., P. J.F., R. P. Bibliographie critique. In: Annuaire français de droit international, volume 14, 1968. pp. 971-1013