Recombinant PDGF from lower vertebrates: receptor binding and immunochemical analysis with metabolically labeled growth factor.
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.
Wang
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Dana-Farber Cancer Institute, Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115.
Shamah
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Stiles
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D016475:3T3 Cells; D000818:Animals; D000918:Antibody Specificity; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009036:Moths; D010982:Platelet-Derived Growth Factor / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013045:Species Specificity; D013724:Teratoma; D014162:Transfection; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis / Q000235:genetics
D007106:Immune Sera; D010982:Platelet-Derived Growth Factor; D011994:Recombinant Proteins; D004247:DNA; D017479:Receptors, Platelet-Derived Growth Factor