Expression of three recombinant homodimeric isoforms of PDGF in Saccharomyces cerevisiae: evidence for difference in receptor binding and functional activities

Growth Factors. 1989;1(3):271-81. doi: 10.3109/08977198908998003.

Abstract

Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A- and B-chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AAS and PDGF-AAL) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AAS and PDGF-AAL were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 nM, respectively. PDGF-BB bound to cells with A- and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A-type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Cell Division / drug effects
  • Fibroblasts / drug effects
  • Genetic Vectors
  • Humans
  • In Vitro Techniques
  • Mitogens
  • Molecular Sequence Data
  • Platelet-Derived Growth Factor / biosynthesis*
  • Platelet-Derived Growth Factor / isolation & purification
  • Platelet-Derived Growth Factor / pharmacology
  • Receptors, Cell Surface / metabolism*
  • Receptors, Platelet-Derived Growth Factor
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology
  • Saccharomyces cerevisiae / metabolism*
  • Transformation, Genetic

Substances

  • Mitogens
  • Platelet-Derived Growth Factor
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Receptors, Platelet-Derived Growth Factor