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

S  CRAIG; J M  CLEMENTS; A L  COOK; D T F  DRYDEN; D R  GREEN; K  HEREMANS; P M  KIRWIN; M J  PRICE; A  FALLON; David Dryden

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

S CRAIG, J M CLEMENTS, A L COOK, D T F DRYDEN, D R GREEN, K HEREMANS, P M KIRWIN, M J PRICE, A FALLON, David Dryden

School of Chemistry

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	67-72
Number of pages	6
Journal	Biochemical Journal
Volume	281
Publication status	Published - 1 Jan 1992

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@article{37df199acfaa414ba5db803deeaad60d,

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.",

author = "S CRAIG and CLEMENTS, \{J M\} and COOK, \{A L\} and DRYDEN, \{D T F\} and GREEN, \{D R\} and K HEREMANS and KIRWIN, \{P M\} and PRICE, \{M J\} and A FALLON and David Dryden",

year = "1992",

month = jan,

day = "1",

language = "English",

volume = "281",

pages = "67--72",

journal = "Biochemical Journal",

issn = "0264-6021",

publisher = "Portland Press Ltd.",

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TY - JOUR

T1 - 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

AU - CRAIG, S

AU - CLEMENTS, J M

AU - COOK, A L

AU - DRYDEN, D T F

AU - GREEN, D R

AU - HEREMANS, K

AU - KIRWIN, P M

AU - PRICE, M J

AU - FALLON, A

AU - Dryden, David

PY - 1992/1/1

Y1 - 1992/1/1

N2 - 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.

AB - 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.

M3 - Article

SN - 0264-6021

VL - 281

SP - 67

EP - 72

JO - Biochemical Journal

JF - Biochemical Journal

ER -

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

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