title: Characterization of the structure and conformation of platelet-derived growth factor-BB (PDGF-BB) and proteinase-resistant mutants of PDGF-BB expressed in Saccharomyces cerevisiae.
abstract: A detailed biophysical study of the secondary and tertiary structures of recombinant platelet-derived growth factor (PDGF)-BB produced in yeast has been carried out. The secondary structure of the molecule is composed of 54% beta-sheet with less than 5% ordered helix. The single tryptophan residue has been shown to be solvent-accessible; however, the ability of the side chain to rotate is severely restricted. The fluorescence emission is quenched at pH 7.0 and in the presence of high salt, but dequenched by titration to lower pH with a pK of 5.8. Two proteinase-resistant mutants of PDGF [( Ser28]- and [Pro32]-PDGF-BB) have also been characterized and shown to have secondary and tertiary structures indistinguishable from wild-type PDGF-BB. These are, therefore, suitable stable background molecules in which to carry out structure-activity-relationship studies on PDGF-BB.
authors.affiliation: British Bio-technology Ltd., Cowley, Oxford, U.K.
authors.forename: S
authors.initials: S
authors.lastname: Craig
authors.forename: J M
authors.initials: JM
authors.lastname: Clements
authors.forename: A L
authors.initials: AL
authors.lastname: Cook
authors.forename: D T
authors.initials: DT
authors.lastname: Dryden
authors.forename: D R
authors.initials: DR
authors.lastname: Green
authors.forename: K
authors.initials: K
authors.lastname: Heremans
authors.forename: P M
authors.initials: PM
authors.lastname: Kirwin
authors.forename: M J
authors.initials: MJ
authors.lastname: Price
authors.forename: A
authors.initials: A
authors.lastname: Fallon
mesh_terms: D016297:Mutagenesis, Site-Directed; D010982:Platelet-Derived Growth Factor / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013059:Spectrum Analysis, Raman
chemical_list: D010982:Platelet-Derived Growth Factor; D011994:Recombinant Proteins