title: The conformation and stability of recombinant-derived granulocyte-macrophage colony stimulating factors.
abstract: The conformation and stability of recombinant-derived human and murine granulocyte-macrophage colony stimulating factors produced in Escherichia coli have been investigated by analytical ultracentrifugation, urea-gradient polyacrylamide gel electrophoresis and several spectroscopic methods. The proteins were demonstrated to be physically homogeneous monomeric proteins with compact globular shapes and shown to have similar secondary structures containing both alpha-helix and beta-sheet structure. The intramolecular disulphide linkages of both proteins were shown to be essential for maintaining native conformation as reduction with dithiothreitol resulted in protein unfolding. Comparison of the human E. coli-derived (non-glycosylated) and mammalian cell culture-derived (glycosylated) proteins by urea-gradient electrophoresis indicated that glycosylation had no major effect on the conformational stability and kinetics of urea induced unfolding and refolding.
authors.affiliation: Glaxo Institute for Molecular Biology, Genève, Switzerland.
authors.forename: P
authors.initials: P
authors.lastname: Wingfield
authors.forename: P
authors.initials: P
authors.lastname: Graber
authors.forename: P
authors.initials: P
authors.lastname: Moonen
authors.forename: S
authors.initials: S
authors.lastname: Craig
authors.forename: R H
authors.initials: RH
authors.lastname: Pain
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003115:Colony-Stimulating Factors / Q000302:isolation & purification; D004220:Disulfides; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014461:Ultracentrifugation
chemical_list: D003115:Colony-Stimulating Factors; D004220:Disulfides; D006133:Growth Substances; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor