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

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 h...

Full description

Saved in:
Bibliographic Details
Published in:Biochemical journal 1992, Vol.281 (1), p.67-72
Main Authors: CRAIG, S, CLEMENTS, J. M, COOK, A. L, DRYDEN, D. T. F, GREEN, D. R, HERAMANS, K, KIRWIN, P. M, PRICE, M. J, FALLON, A
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
conformation
fluorescence
Fundamental and applied biological sciences. Psychology
Mutagenesis, Site-Directed
mutants
platelet-derived growth factor
Platelet-Derived Growth Factor - chemistry
Platelet-Derived Growth Factor - genetics
Platelet-Derived Growth Factor - isolation & purification
Protein Conformation
Protein hormones. Growth factors. Cytokines
proteinase
Proteins
Recombinant Proteins - chemistry
Recombinant Proteins - isolation & purification
resistant
Saccharomyces cerevisiae - genetics
Spectrometry, Fluorescence
Spectrophotometry, Ultraviolet
Spectrum Analysis, Raman
structure
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj2810067