title: Probing the interaction of brain fatty acid binding protein (B-FABP) with model membranes.
abstract: Brain fatty acid-binding protein (B-FABP) interacts with biological membranes and delivers polyunsaturated fatty acids (FAs) via a collisional mechanism. The binding of FAs in the protein and the interaction with membranes involve a motif called "portal region", formed by two small α-helices, A1 and A2, connected by a loop. We used a combination of site-directed mutagenesis and electron spin resonance to probe the changes in the protein and in the membrane model induced by their interaction. Spin labeled B-FABP mutants and lipidic spin probes incorporated into a membrane model confirmed that B-FABP interacts with micelles through the portal region and led to structural changes in the protein as well in the micelles. These changes were greater in the presence of LPG when compared to the LPC models. ESR spectra of B-FABP labeled mutants showed the presence of two groups of residues that responded to the presence of micelles in opposite ways. In the presence of lysophospholipids, group I of residues, whose side chains point outwards from the contact region between the helices, had their mobility decreased in an environment of lower polarity when compared to the same residues in solution. The second group, composed by residues with side chains situated at the interface between the α-helices, experienced an increase in mobility in the presence of the model membranes. These modifications in the ESR spectra of B-FABP mutants are compatible with a less ordered structure of the portal region inner residues (group II) that is likely to facilitate the delivery of FAs to target membranes. On the other hand, residues in group I and micelle components have their mobilities decreased probably as a result of the formation of a collisional complex. Our results bring new insights for the understanding of the gating and delivery mechanisms of FABPs.
authors.affiliation: Grupo de Biofísica Molecular Sérgio Mascarenhas, Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, Sao Paulo, Brazil.
authors.forename: Fábio
authors.initials: F
authors.lastname: Dyszy
authors.forename: Andressa P A
authors.initials: AP
authors.lastname: Pinto
authors.forename: Ana P U
authors.initials: AP
authors.lastname: Araújo
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Costa-Filho
mesh_terms: D001921:Brain / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D008567:Membranes, Artificial; D008823:Micelles; D016297:Mutagenesis, Site-Directed
chemical_list: D050556:Fatty Acid-Binding Proteins; D005231:Fatty Acids, Unsaturated; D008567:Membranes, Artificial; D008823:Micelles
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