Identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells
The binding of biologically active, SVI-labeled basic fibroblast growth factor (FGF) to baby hamster kidney-derived cell line cells (BHK-21) was studied at 4 degrees C. Unlabeled FGF displaced cell surface bound SVI-FGF, but platelet-derived growth factor, epidermal growth factor, insulin, or transferrin did not. Binding was saturable both as a function of time and as a function of increasing SVI-FGF concentrations. Scatchard analysis of the binding data revealed the presence of about 1.2 X 10(5) binding sites/cell with an apparent KD of 270 pM. The number of the binding sites was down-regulated following preincubation of the cells with FGF. The density of binding sites/cell also decreased as an inverse function of cell density. When SVI-FGF binding was studied in a BHK-21 cell membrane preparation, it was found that the membranal binding site displayed a lower KD of 21 pM. SVI-FGF was covalently cross-linked to its cell surface receptor on intact BHK-21 cells using the homobifunctional agent disuccinimidyl suberate. Two macromolecular species with an apparent molecular weight of 145,000 and SV,000, respectively, were labeled under both reducing and nonreducing conditions. Unlabeled FGF competed with SVI-FGF for binding to both macromolecular species. The labeling of the macromolecules was also inhibited by heparin. No labeling was observed in the absence of the cross-linkers or when heat-inactivated SVI-FGF was used instead of radiolabeled, biologically active FGF.
- Research Organization:
- Univ. of California Medical Center, San Francisco
- OSTI ID:
- 6336053
- Journal Information:
- J. Biol. Chem.; (United States), Vol. 25
- Country of Publication:
- United States
- Language:
- English
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HORMONES
BIOCHEMICAL REACTION KINETICS
RECEPTORS
CELL MEMBRANES
COVALENCE
CROSS-LINKING
FIBROBLASTS
HAMSTERS
HEPARIN
INSULIN
IODINE 125
KIDNEYS
LIGANDS
MOLECULAR WEIGHT
PHOSPHORYLATION
TRACER TECHNIQUES
TRANSFERRIN
AMINES
ANIMAL CELLS
ANIMALS
ANTICOAGULANTS
BETA DECAY RADIOISOTOPES
BODY
CARBOHYDRATES
CELL CONSTITUENTS
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
GLOBULINS
GLOBULINS-BETA
HEMATOLOGIC AGENTS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
MAMMALS
MEMBRANES
METALLOPROTEINS
MUCOPOLYSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PEPTIDE HORMONES
POLYMERIZATION
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SACCHARIDES
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques