Recombinant PDGF from lower vertebrates: receptor binding and immunochemical analysis with metabolically labeled growth factor

Growth Factors. 1992;7(4):279-88. doi: 10.3109/08977199209046410.

Abstract

We used a baculovirus vector/insect host cell system to express cDNA clones of PDGF A genes from mouse and frog (Xenopus laevis). The insect host cells process PDGF A subunits from either frogs or mice into biologically active AA homodimers with yields in the range of 0.5-1.0 mg/liter of culture medium. The recombinant PDGFs can be metabolically labeled with 35S-cysteine for use in radioreceptor and radioimmunoassays. Neutralizing polyclonal antisera can be raised against the mouse and frog PDGFs. These antisera are markedly species-specific in action. However, in radioreceptor binding assays and bioassays for mitogenic activity, human, mouse and frog PDGF AA homodimers occupy and activate murine PDGF receptors with equal efficiency.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • Antibody Specificity
  • Baculoviridae / genetics
  • Cell Line
  • DNA / genetics
  • Gene Expression
  • Humans
  • Immune Sera / immunology
  • Mice
  • Mice, Inbred BALB C
  • Moths
  • Platelet-Derived Growth Factor / genetics
  • Platelet-Derived Growth Factor / immunology
  • Platelet-Derived Growth Factor / metabolism*
  • Receptors, Platelet-Derived Growth Factor / metabolism*
  • Recombinant Proteins / metabolism
  • Species Specificity
  • Teratoma
  • Transfection
  • Tumor Cells, Cultured
  • Xenopus laevis / genetics

Substances

  • Immune Sera
  • Platelet-Derived Growth Factor
  • Recombinant Proteins
  • DNA
  • Receptors, Platelet-Derived Growth Factor