Fibroblast growth factor binding to perlecan isolated  from the growth plate

Abstract 2005 MHE Conference

Leigh Westa, Prasanthi Govindraja, Xiuqin Zhangb, David M. Ornitz(b), John R. Hassell, (a)Center for Research in
Skeletal Development and  Pediatric Orthopaedics,
Shriners Hospitals for Children, Tampa, FL 33612   
(b)Department of Molecular Biology and Pharmacology, Washington  University Medical School,660 S. Euclid
Avenue, St. Louis, MO 63310,
(c)Department of Biochemistry and Molecular Biology,College of  Medicine, University of  South Florida, Tampa, FL
33612

Fibroblast growth factor (FGF)-2, FGF-9 and FGF-18 are three FGF's that have been shown to regulate chondrocyte proliferation
in the growth plate.  

Heparin and heparan sulfate proteoglycans from endothelial cells have been shown to bind FGF-2. Perlecan is present in the
growth plate as a proteoglycan containing both heparan and chondroitin sulfate chains and the phenotype of the perlecan
knockout mouse shows it is necessary for the proliferation of chondrocytes in the growth plate.

We evaluated the binding of FGF's to perlecan from the growth plate using a cationic filtration assay.  We found FGF-2 bound
primary to the heparan sulfate chains on perlecan but that the core protein was also involved in FGF-2 binding when the
heparan sulfate chains were present.  Removal of Chondroitin sulfate chains on perlecan enhanced FGF-2 binding slightly.  
FGF-1, FGF-9 and FGF-18 showed no binding to perlecan but FGF-7 showed a low level of binding.  

We also found, using recombinant FGF receptors in a separate capture assay that while the receptors would bind to a
heparin-FGF-2 complex, the receptors would not bind to the perlecan-FGF-2 complex.  Similarly, perlecan did not augment
FGF-2 stimulation of [3H]-thymine incorporation in BaF3 cells.   

These data show perlecan can bind to FGF-2 by its heparan sulfate chains but that chondroitin sulfate chains on the perlecan
acts to block transfer of the bound FGF-2 to the receptor.
Research authored by Dr. Hassell
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