title: High-pressure effects on beta-lactoglobulin interactions with ligands studied by fluorescence.
abstract: The effects of pressure (0.1 MPa to 400 MPa) on intrinsic fluorescence of beta-lactoglobulin and on its binding of retinol and cis-parinaric acid have been studied at neutral and acid pHs. In neutral pH, fluorescence emission spectra of beta-lactoglobulin tryptophanes are characterized by an irreversible 14 nm red-shift indicating pressure-induced folding changes. The intensity of the fluorescence of retinol in beta-lactoglobulin-retinol complex is enhanced by a pressure increase up to 150 MPa. It decreases at higher pressures and disappears altogether at 300 MPa. beta-Lactoglobulin-retinol complex does not reassociate after decompression at neutral pH. At acid pH condition, the fluorescence quenching by pressure of beta-lactoglobulin tryptophans is coupled with a 2 nm spectral shift and is fully reversible demonstrating almost complete restoration of globulin folding. The evolution of retinol fluorescence in beta-lactoglobulin-retinol complex is also entirely reversible between 0.1 MPa and 400 MPa and the complex never dissociates in the studied pressure range. beta-lactoglobulin-cis-parinaric acid complexes at neutral and acid pH values dissociate irreversibly at 200 MPa and 350 MPa, respectively.
authors.affiliation: L.E.I.M.A. Institut National de la Recherche Agronomique, Nantes, France.
authors.forename: E
authors.initials: E
authors.lastname: Dufour
authors.forename: G H
authors.initials: GH
authors.lastname: Hoa
authors.forename: T
authors.initials: T
authors.lastname: Haertlé
mesh_terms: D001665:Binding Sites; D005231:Fatty Acids, Unsaturated / Q000737:chemistry*; D005990:Glycerol / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry*; D008024:Ligands; D011312:Pressure; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D014801:Vitamin A / Q000737:chemistry*
chemical_list: D005231:Fatty Acids, Unsaturated; D007782:Lactoglobulins; D008024:Ligands; D014801:Vitamin A; D005990:Glycerol; C013641:parinaric acid