RRC ID 61645
Author Terada M, Shimizu A, Sato N, Miyakaze SI, Katayama H, Kurokawa-Seo M.
Title Fibroblast growth factor receptor 3 lacking the Ig IIIb and transmembrane domains secreted from human squamous cell carcinoma DJM-1 binds to FGFs.
Journal Mol Cell Biol Res Commun
Abstract The fibroblast growth factor receptors (FGFRs) are a family of transmembrane tyrosine kinases that play a key role in cell growth and tumorigenesis in response to FGFs. FGFR complexity is increased by the existence of additional isoforms generated by alternative mRNA splicing. We identified that the transcript FGFR3DeltaTM, an alternatively spliced isoform of FGFR3 lacking exons encoding the C-terminal half of Ig III (IIIb) and transmembrane domains, is expressed in the human squamous carcinoma cell line DJM-1. To determine whether FGFR3DeltaTM has the potential to be secreted, we analyzed the protein expression in CHOK1 cells transfected with FGFR3DeltaTM cDNA and DJM-1 cells. Western blot analysis revealed that FGFR3DeltaTM protein was secreted, N-glycosylated, and dimerized by an intermolecular disulfide bond. Cross-linking experiments showed that FGF1 and FGF2 were able to bind to FGFR3DeltaTM, suggesting that the loss of the Ig IIIb domain may confer upon FGFR3DeltaTM the ability to bind to FGF2.
Volume 4(6)
Pages 365-73
Published 2001-11-1
DOI 10.1006/mcbr.2001.0306
PII S152247240190306X
PMID 11703096
MeSH Alternative Splicing Amino Acid Sequence Animals Base Sequence Blotting, Western CHO Cells Carcinoma, Squamous Cell / genetics Carcinoma, Squamous Cell / metabolism Cell Membrane / metabolism Cells, Cultured Cloning, Molecular Cricetinae Cross-Linking Reagents / pharmacology DNA, Complementary / metabolism Disulfides Exons Glycoside Hydrolases / pharmacology Glycosylation Heparin / metabolism Humans Models, Biological Molecular Sequence Data Precipitin Tests Protein Binding Protein Isoforms Protein Structure, Tertiary Protein-Tyrosine Kinases* RNA / metabolism Receptor, Fibroblast Growth Factor, Type 3 Receptors, Fibroblast Growth Factor / chemistry* Receptors, Fibroblast Growth Factor / genetics Receptors, Fibroblast Growth Factor / metabolism* Recombinant Proteins / metabolism Reverse Transcriptase Polymerase Chain Reaction Sepharose / pharmacology Transfection Tumor Cells, Cultured
Human and Animal Cells HBL-100(RCB0460) CHO-K1(RCB0285)