title: Unfolding and refolding of hen egg-white riboflavin binding protein.
abstract: The unfolding and refolding of riboflavin-binding protein (RfBP) from hen egg-white induced by addition of guanidinium chloride (GdnHCl), and its subsequent removal by dialysis have been studied by c.d. and fluorescence for both the native and reduced protein. The reduction of its nine disulphide bonds causes a reduction in the secondary structure (alpha-helix plus beta-sheet) from 63% to 33% of the amino acid residues. Unfolding of the native protein occurred in two phases; the first involving a substantial loss of tertiary structure, followed by a second phase involving loss of secondary structure at higher GdnHCl concentrations. By contrast this biphasic behaviour was not discernible in the reduced protein. The loss of ability to bind riboflavin occurred after the first phase of unfolding. Comparison of unfolding of the holoprotein and apoprotein suggested that riboflavin has only a small stabilizing effect on the unfolding process. After removal of GdnHCl, the holoprotein, apoprotein and reduced protein assumed their original conformation. The significance of the results in relation to various models for protein folding is discussed.
authors: Department of Biological and Molecular Sciences, Stirling University, UK.
authors: S
authors: S
authors: Allen
authors: L
authors: L
authors: Stevens
authors: D
authors: D
authors: Duncan
authors: S M
authors: SM
authors: Kelly
authors: N C
authors: NC
authors: Price
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002645:Chickens; D002942:Circular Dichroism; D004220:Disulfides / Q000737:chemistry; D004527:Egg Proteins / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D026901:Membrane Transport Proteins; D010084:Oxidation-Reduction; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012256:Riboflavin / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D002352:Carrier Proteins; D004220:Disulfides; D004527:Egg Proteins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein; D012256:Riboflavin