title: Structural and conformational changes of beta-lactoglobulin B: an infrared spectroscopic study of the effect of pH and temperature.
abstract: Infrared spectra of 2.5 mM solutions of beta-lactoglobulin B were recorded as a function of pH (from pH 2 to pH 13) and as a function of temperature (from -100 degrees C to +90 degrees C). An analysis of the pH- and temperature-induced changes in the secondary structure was performed based on changes in the conformation-sensitive amide I bands of beta-lactoglobulin. Whereas the total amount of beta-structure remains constant (56-59%) between pH 2 and pH 10, the proportions of the various beta-components do change. In particular, the dimerization of the monomeric protein, induced by raising the pH from 2 to 3 , leads to an increase in the intensity of the 1636 cm-1 band (associated with antiparallel beta-sheet), at the expense of the 1626 cm-1 band (associated with exposed beta-strands). Both the thermal and alkaline denaturation of beta-lactoglobulin occur in two distinct stages. Although the spectra (i.e., the structures) after complete thermal or alkaline denaturation are clearly different, the spectrum of the protein after the first stage of thermal denaturation (at about 60 degrees C) is the same as that after the first stage of alkaline denaturation (at pH 11), suggesting a common denaturation intermediate, which probably represents a crossover point in a complex potential hypersurface.
authors.affiliation: Division of Chemistry, National Research Council of Canada, Ottawa.
authors.forename: H L
authors.initials: HL
authors.lastname: Casal
authors.forename: U
authors.initials: U
authors.lastname: Köhler
authors.forename: H H
authors.initials: HH
authors.lastname: Mantsch
mesh_terms: D000577:Amides; D000818:Animals; D002417:Cattle; D005990:Glycerol / Q000494:pharmacology; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D013055:Spectrophotometry, Infrared; D013696:Temperature
chemical_list: D000577:Amides; D007782:Lactoglobulins; D005990:Glycerol