title: The integrity of the alpha-helical domain of intestinal fatty acid binding protein is essential for the collision-mediated transfer of fatty acids to phospholipid membranes.
abstract: Intestinal FABP (IFABP) and liver FABP (LFABP), homologous proteins expressed at high levels in intestinal absorptive cells, employ markedly different mechanisms of fatty acid transfer to acceptor model membranes. Transfer from IFABP occurs during protein-membrane collisional interactions, while for LFABP transfer occurs by diffusion through the aqueous phase. In addition, transfer from IFABP is markedly faster than from LFABP. The overall goal of this study was to further explore the structural differences between IFABP and LFABP which underlie their large functional differences in ligand transport. In particular, we addressed the role of the alphaI-helix domain in the unique transport properties of intestinal FABP. A chimeric protein was engineered with the 'body' (ligand binding domain) of IFABP and the alphaI-helix of LFABP (alpha(I)LbetaIFABP), and the fatty acid transfer properties of the chimeric FABP were examined using a fluorescence resonance energy transfer assay. The results showed a significant decrease in the absolute rate of FA transfer from alpha(I)LbetaIFABP compared to IFABP. The results indicate that the alphaI-helix is crucial for IFABP collisional FA transfer, and further indicate the participation of the alphaII-helix in the formation of a protein-membrane "collisional complex". Photo-crosslinking experiments with a photoactivable reagent demonstrated the direct interaction of IFABP with membranes and further support the importance of the alphaI helix of IFABP in its physical interaction with membranes.
authors.affiliation: Instituto de Investigaciones Bioquímicas de La Plata, CONICET-UNLP, Facultad de Ciencias Médicas, calles 60 y 120, 1900-La Plata, Argentina.
authors.forename: G R
authors.initials: GR
authors.lastname: Franchini
authors.forename: J
authors.initials: J
authors.lastname: Storch
authors.forename: B
authors.initials: B
authors.lastname: Corsico
mesh_terms: D000818:Animals; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D008563:Membrane Lipids / Q000378:metabolism*; D008956:Models, Chemical; D010743:Phospholipids / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D053835:Unilamellar Liposomes / Q000378:metabolism
chemical_list: C493321:Fabp1 protein, rat; C493407:Fabp2 protein, rat; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D008563:Membrane Lipids; D010743:Phospholipids; D011993:Recombinant Fusion Proteins; D053835:Unilamellar Liposomes