title: Circular dichroism of reduced and oxidized recombinant human epidermal growth factor.
abstract: To further elucidate the role of the disulfide bonds in determining the protein folding of recombinant human epidermal growth factor (r-HuEGF) we studied the structure of reduced and oxidized r-HuEGF using circular dichroism (CD). The far UV CD spectrum of reduced r-HuEGF in 10 mM sodium phosphate pH 3.0 is very different from that of the oxidized molecule. The spectrum of the reduced molecule consists of a plateau from 225 to 200 nm, consistent with the presence of alpha-helix, beta-sheet, and unordered structure. The addition of the alpha-helix inducer trifluoroethanol to the reduced molecule resulted in an enhancement of alpha-helix, at the apparent expense of beta-sheet, while the oxidized molecule was unaffected by the presence of this reagent. Secondary structure predictions based on the amino acid sequence of EGF correlate most closely with the structure of the reduced molecule. From these results, it appears that the r-HuEGF has a more regular secondary structure in the absence of the disulfide bonds than in their presence. This suggests that the folding of EGF occurs by destroying the regular secondary structure that was present in the reduced state, and that the structure of the native molecule is dictated largely by disulfide bonding.
authors.affiliation: Amgen Inc., Amgen Center, Thousand Oaks, CA.
authors.forename: L O
authors.initials: LO
authors.lastname: Narhi
authors.forename: T
authors.initials: T
authors.lastname: Arakawa
authors.forename: M D
authors.initials: MD
authors.lastname: McGinley
authors.forename: M F
authors.initials: MF
authors.lastname: Rohde
authors.forename: K R
authors.initials: KR
authors.lastname: Westcott
mesh_terms: D002942:Circular Dichroism; D004220:Disulfides / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000737:chemistry*; D006801:Humans; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor