Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts.

2.50
Hdl Handle:
http://hdl.handle.net/10541/83573
Title:
Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts.
Authors:
Delehedde, Maryse; Lyon, Malcolm; Gallagher, John T; Rudland, Philip S; Fernig, David G
Abstract:
We examine the relationship between the chain length of heparin-derived oligosaccharides, fibroblast growth factor (FGF)-2 binding 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 tetrasaccharides. As the chain length increased from tetrasaccharide to octasaccharide, there was a substantial increase in k(ass) (564000 M(-1) x s(-1) to 2000000 M(-1) x 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) x 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.
Affiliation:
School of Biological Sciences, Life Science Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
Citation:
Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts. 2002, 366 (Pt 1):235-44 Biochem. J.
Journal:
The Biochemical Journal
Issue Date:
15-Aug-2002
URI:
http://hdl.handle.net/10541/83573
DOI:
10.1042/BJ20011718
PubMed ID:
12000311
Type:
Article
Language:
en
ISSN:
0264-6021
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorDelehedde, Maryse-
dc.contributor.authorLyon, Malcolm-
dc.contributor.authorGallagher, John T-
dc.contributor.authorRudland, Philip S-
dc.contributor.authorFernig, David G-
dc.date.accessioned2009-10-05T15:18:29Z-
dc.date.available2009-10-05T15:18:29Z-
dc.date.issued2002-08-15-
dc.identifier.citationFibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts. 2002, 366 (Pt 1):235-44 Biochem. J.en
dc.identifier.issn0264-6021-
dc.identifier.pmid12000311-
dc.identifier.doi10.1042/BJ20011718-
dc.identifier.urihttp://hdl.handle.net/10541/83573-
dc.description.abstractWe examine the relationship between the chain length of heparin-derived oligosaccharides, fibroblast growth factor (FGF)-2 binding 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 tetrasaccharides. As the chain length increased from tetrasaccharide to octasaccharide, there was a substantial increase in k(ass) (564000 M(-1) x s(-1) to 2000000 M(-1) x 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) x 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.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshBinding, Competitive-
dc.subject.meshBiotinylation-
dc.subject.meshBlotting, Western-
dc.subject.meshCell Division-
dc.subject.meshDNA-
dc.subject.meshDimerization-
dc.subject.meshDose-Response Relationship, Drug-
dc.subject.meshFibroblast Growth Factor 2-
dc.subject.meshFibroblasts-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshHeparin-
dc.subject.meshHumans-
dc.subject.meshKinetics-
dc.subject.meshMitogen-Activated Protein Kinase 1-
dc.subject.meshMitogen-Activated Protein Kinase 3-
dc.subject.meshMitogen-Activated Protein Kinases-
dc.subject.meshOligosaccharides-
dc.subject.meshPhosphorylation-
dc.subject.meshPolysaccharides-
dc.subject.meshProtein Binding-
dc.subject.meshRats-
dc.subject.meshRecombinant Proteins-
dc.subject.meshRibosomal Protein S6 Kinases-
dc.subject.meshTime Factors-
dc.titleFibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts.en
dc.typeArticleen
dc.contributor.departmentSchool of Biological Sciences, Life Science Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.en
dc.identifier.journalThe Biochemical Journalen

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