title: Alcohol-induced changes of beta-lactoglobulin-retinol-binding stoichiometry.
abstract: It has been demonstrated using CD that ethanol induces important secondary structure changes of beta-lactoglobulin. CD spectra indicate that beta-lactoglobulin secondary structure, which is mainly composed of beta-strands, becomes mostly alpha-helical under the influence of the solvent polarity changes. The midpoint of beta-strand/alpha-helix transition in beta-lactoglobulin is observed at dielectric constant approximately 60 (35% ethanol; v/v). According to CD measurements, the ethanol-dependent secondary structure changes are reversible. The alkylation of lysines epsilon-NH2 in beta-lactoglobulin weakens the central beta-barrel structure, since the beta-strand/alpha-helix transition midpoint of alkylated beta-lactoglobulin is shifted to lower ethanol concentration (25% ethanol; v/v). beta-Lactoglobulin structural changes are triggering the dissociation of the beta-lactoglobulin-retinol complex as judged from complete quenching of its fluorescence in ethanol concentration greater than 30% (v/v). However, in 20% ethanol (v/v), beta-lactoglobulin still retains most of its native secondary structure as shown by CD and, in this condition, one beta-lactoglobulin molecule binds an additional second retinol molecule. This suggests that the highly populated species observed around 20% ethanol (v/v) might represent an intermediate state able to bind two molecules of retinol.
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: T
authors.initials: T
authors.lastname: Haertlé
mesh_terms: D000478:Alkylation; D002942:Circular Dichroism; D000431:Ethanol / Q000494:pharmacology*; D005453:Fluorescence; D007782:Lactoglobulins / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D014801:Vitamin A / Q000737:chemistry* / Q000378:metabolism
chemical_list: D007782:Lactoglobulins; D014801:Vitamin A; D000431:Ethanol