Journal
BIOCHEMICAL JOURNAL
Volume 366, Issue -, Pages 235-244Publisher
PORTLAND PRESS LTD
DOI: 10.1042/BJ20011718
Keywords
binding kinetics; fibroblast growth factor receptor; glycosaminoglycan; heparan sulphate; optical biosensor
Categories
Ask authors/readers for more resources
We examine the relationship between the chain length of heparin-derived oligosaccharides, fibroblast growth factor (FGF)-2 bindin g kinetics and the ability of the oligosaccharides to allow FGF-2-induced proliferation of chlorate-treated rat mammary fibroblasts. First, using an optical biosensor, we show that FGF-2 did not bind disaccharides, but definitively bound to tetra-saccharides. As the chain length increased from tetrasaccharide to octasaccharide, there was a substantial increase in k(ass) (564000 M-1 . s(-1) to 2000000 M-1 . s-1, respectively) and affinity (K-d 77 nM to 11 nM, respectively) for FGF-2. From decasaccharides and longer, the k(ass) and affinity for FGF-2) was reduced substantially (tetradecasaccharide k(ass) 470000 M-1 . s(-1), K-d 30 nM). In chlorate-treated, and hence sulphated, glycosaminoglycan-deficient cells, FGF-2 alone or in the presence of disaccharides did not stimulate DNA synthesis and it only elicited an early transient dual phosphorylation of p42/44 mitogen-activated protein kinase (MAPK). In the same cells FGF-2 in the presence of tetrasaccharides and longer oligosaccharides was able to restore DNA synthesis and enable the sustained dual phosphorylation of p42/44(MAPK). However, the oligosaccharides from tetrasaccharides to octasaccharides were less potent in proliferation assays than deca- and longer oligosaccharides, Therefore, there was no correlation between the binding parameters and the potency of the oligosaccharides in DNA synthesis assays. These results demonstrate that tetrasaccharides are able to bind FGF-2 and enable FGF-2 to stimulate cell proliferation, which sets important boundary conditions for models of the FGF-2-heparan sulphate-FGF receptor complex.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available