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

被引:5
作者
CRAIG, S
CLEMENTS, JM
COOK, AL
DRYDEN, DTF
GREEN, DR
HEREMANS, K
KIRWIN, PM
PRICE, MJ
FALLON, A
机构
[1] UNIV NEWCASTLE UPON TYNE,SCH MED,DEPT BIOCHEM & GENET,NEWCASTLE TYNE NE2 4HH,ENGLAND
[2] KATHOLIEKE UNIV LEUVEN,DEPT CHEM,CHEM & BIOL DYNAM LAB,B-3001 LOUVAIN,BELGIUM
关键词
D O I
10.1042/bj2810067
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:67 / 72
页数:6
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