A catalytic role of heparin within the extracellular matrix.
We investigated the mechanism by which heparin enhances the binding of vascular endothelial growth factor (VEGF) to the extracellular matrix protein fibronectin. In contrast to other systems, where heparin acts as a protein scaffold, we found that heparin functions catalytically to modulate VEGF binding site availability on fibronectin. By measuring the binding of VEGF and heparin to surface-immobilized fibronectin, we show that substoichiometric amounts of heparin exposed cryptic VEGF binding sites within fibronectin that remain available after heparin removal. Measurement of association and dissociation kinetics for heparin binding to fibronectin indicated that the interaction is rapid and transient. We localized the heparin-responsive element to the C-terminal 40-kDa Hep2 domain of fibronectin. A mathematical model of this catalytic process was constructed that supports a mechanism whereby the heparin-induced conformational change in fibronectin is accompanied by release of heparin. Experiments with endothelial extracellular matrix suggest that this process may also occur within biological matrices. These results indicate a novel mechanism whereby heparin catalyzes the conversion of fibronectin to an open conformation by transiently interacting with fibronectin and progressively hopping from molecule to molecule. Catalytic activation of the extracellular matrix might be an important mechanism for heparin to regulate function during normal and disease states.
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Maria
M
Mitsi
Kimberly
K
Forsten-Williams
Manoj
M
Gopalakrishnan
Matthew A
MA
Nugent
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry; D006493:Heparin / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry
D000925:Anticoagulants; D005353:Fibronectins; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D006493:Heparin