Requirement for anticoagulant heparan sulfate in the fibroblast growth factor receptor complex.
A divalent cation-dependent association between heparin or heparan sulfate and the ectodomain of the fibroblast growth factor (FGF) receptor kinase (FGFR) restricts FGF-independent trans-phosphorylation between self-associated FGFR and determines specificity for and mediates binding of activating FGF. Here we show that only the fraction of commercial heparin or rat liver heparan sulfate which binds to immobilized antithrombin formed an FGF-binding binary complex with the ectodomain of the FGFR kinase. Conversely, only the fraction of heparin that binds to immobilized FGFR inhibited Factor Xa in the presence of antithrombin. Only the antithrombin-bound fraction of heparin competed with (3)H-heparin bound to FGFR in absence of FGF, whereas both antithrombin-bound and unretained fractions competed with radiolabeled heparin bound independently to FGF-1 and FGF-2. The antithrombin-bound fraction of heparin was required to support the heparin-dependent stimulation of DNA synthesis of endothelial cells by FGF-1. The requirement for divalent cations and the antithrombin-binding motif distinguish the role of heparan sulfate as an integral subunit of the FGFR complex from the wider range of effects of heparan sulfates and homologues on FGF signaling through FGFR-independent interactions with FGF.
Department of Biochemistry and Biophysics, Texas A&M University, Texas A&M University System Health Science Center, Houston, Texas 77030-3303, USA.
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D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification / Q000378:metabolism; D000991:Antithrombins / Q000378:metabolism; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D016220:Fibroblast Growth Factor 1; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans / Q000302:isolation & purification / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000302:isolation & purification / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000302:isolation & purification / Q000378:metabolism*; D013552:Swine
D000925:Anticoagulants; D000991:Antithrombins; D016220:Fibroblast Growth Factor 1; D016222:Fibroblast Growth Factor 2; D005346:Fibroblast Growth Factors; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin; D006497:Heparan Sulfate; D017468:Receptors, Fibroblast Growth Factor