Molecular cloning of a human basic fibroblast growth factor receptor cDNA and expression of a biologically active extracellular domain in a baculovirus system

Michael C. Kiefer, Andrew Baird, Tuan Nguyen, Carlos George-Nascimento, Owen B. Mason, Leslie J. Boley, Pablo Valenzuela, Philip J. Barr

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

33 Citas (Scopus)

Resumen

A cDNA clone encoding a human fibroblast growth factor (FGF) receptor was isolated from a hepatoma cell line cDNA library. The cDNA encodes a three immunoglobulinlike-domain FGF receptor that is similar to a human placental FGF receptor cDNA but lacks two amino acids. The variation observed at these two amino acids, also seen in the two immunoglobulinlike-domain FGF-receptors, can be explained by an alternate splicing mechanism. We have used a baculovirus expression system to produce high levels of a soluble, extracellular domain form of the FGF receptor (EC-FGF receptor). Spodoptera frugiperda (Sf9) insect cells infected with recombinant EC-FGF receptor viruses synthesized and secreted an EC-FGF receptor of apparent Mt=58,000. The EC-FGF receptor purified from conditioned media of infected Sf9 cells by lentil lectin affinity chromatography was shown to bind basic FGF with high affinity (Kd= 1-5 nM), to inhibit the binding of radioiodinated basic FGF to its high affinity receptor and to inhibit endothelial cell proliferation. Furthermore, binding of basic FGF to the EC-FGF receptor was shown to be significantly enhanced by heparin. The availability of biologically active FGF receptors will allow an analysis of their interaction with members of the FGF family of proteins and viruses of the herpes family that have been shown to use the FGF receptor system for cell entry.

Idioma originalInglés
Páginas (desde-hasta)115-127
Número de páginas13
PublicaciónGrowth Factors
Volumen5
N.º2
DOI
EstadoPublicada - 1 ene. 1991

Áreas temáticas de ASJC Scopus

  • Endocrinología
  • Bioquímica clínica
  • Biología celular

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