Binding of heparin by type III domains and peptides from the carboxy terminal Hep-2 region of fibronectin

Ingham, Kenneth C.; Brew, Shelesa A.; Migliorini, Mary; Busby, Thomas F.

doi:10.1021/bi00097a035

Cited by 47 publications

(36 citation statements)

References 26 publications

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“…The loop connecting strands C-C' would lay on the GFCC'C" face of the AWN model while the N-terminal stretch might not be constraint in a fixed conformation. It is noteworthy that similar to our results concerning AWN 4RRSR8, the RRAR sequence, when present in a synthetic peptide, has been shown to have weak affinity for heparin [28,29]. Furthermore, this same RRAR sequence exists in the carboxylterminal lobe of lactoferrin where it is not functional as a GAG-binding site [30].…”

Section: Discussionsupporting

confidence: 87%

Mapping the heparin‐binding domain of boar spermadhesins

et al. 1996

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Boar spermadhesins are a group of seminal plasma, heparin-binding proteins which appear to be involved in sperm capacitation and gamete interaction. Using a proteolytic protection assay we have identified regions of AQN-I, AQN-3, PSP-I and AWN which remain attached to a heparin-Sepharose column following in-column digestion of bound spermadhesins with chymotrypsin and elastase. In addition, the complete amino acid sequence of spermadhesin AWN was synthesized as overlapping peptides, and their ability to bind to a heparin-Sepharose column and to inhibit the interaction of soluble heparin with purified ELISA plate-coated AWN was tested. Both approaches gave similar results and as a whole showed that different regions of AWN may converge in its tertiary structure to form a composite heparin-binding site. The conformational heparin-binding surface resides on the GFCC'C" face of the proposed structural model for AWN and is in an opposite location to the carbohydratebinding region of the spermadhesin.

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Section: Discussionsupporting

confidence: 87%

Mapping the heparin‐binding domain of boar spermadhesins

et al. 1996

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“…This is supported by studies where the use of heparinbinding synthetic peptides led to a reduction of affinity (28,29). As a matter of fact, HepV was shown to fold into the characteristic collagenous structure, the polyproline II helix, by circular dichroism, whereas the peptide tested in the same experimental condition was not (data not shown).…”

Section: Discussionmentioning

confidence: 67%

Molecular Features of the Collagen V Heparin Binding Site

Delacoux¹,

Fichard

Geourjon

et al. 1998

Journal of Biological Chemistry

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A heparin binding region is known to be present within the triple helical part of the ␣1(V) chain. Here we show that a recombinant ␣1(V) fragment (Ile 824 to Pro 950 ), referred to as HepV, is sufficient for heparin binding at physiological ionic strength. Both native individual ␣1(V) chains and HepV are eluted at identical NaCl concentrations (0.35 M) from a heparin-Sepharose column, and this binding can be inhibited specifically by the addition of free heparin or heparan sulfate. In contrast, a shorter 23-residue synthetic peptide, containing the putative heparin binding site in HepV, fails to bind heparin. Interestingly, HepV promotes cell attachment, and HepV-mediated adhesion is inhibited specifically by heparin or heparan sulfate, indicating that this region might behave as an adhesive binding site. The same site is equally functional on triple helical molecules as shown by heparin-gold labeling. However, the affinities for heparin of each of the collagen V molecular forms tested are different and increase with the number of ␣1(V) chains incorporated in the molecules. Molecular modeling of a sequence encompassing the putative HepV binding sequence region shows that all of the basic residues cluster on one side of the helical face. A highly positively charged ring around the molecule is thus particularly evident for the ␣1(V) homotrimer. This could strengthen its interaction with the anionic heparin molecules. We propose that a single heparin binding site is involved in heparin-related glycosaminoglycanscollagen V interactions, but the different affinities observed likely modulate cell and matrix interactions between collagen V and heparan sulfate proteoglycans in tissues.Collagen V is a fibrillar collagen that plays an important role in fibrillogenesis, and it also acts as an adhesive substrate for a large variety of cells and binds to a number of extracellular components through its major triple helical domain (1). Collagen V interacts with matrix proteoglycans such as the two small proteoglycans decorin and biglycan (2), the proteoglycan form of macrophage colony-stimulating factor (3), the cell surface proteoglycan syndecan-1 (4, 5), and as shown recently, the membrane spanning proteoglycan NG2 (6). Some of these interactions are mediated by the core proteins, but others depend on the glycosaminoglycan chains such as the heparan sulfate chains.Apart from in vitro binding of collagen V to membranespanning proteoglycans, the suggestion that heparan sulfate interacts with collagen V was supported by inhibition experiments showing a reduction of cell attachment to collagen V in the presence of heparin (7). It has been shown already that cell focal adhesion on fibronectin requires the cooperation of both cell transmembrane proteoglycans and integrin receptors (8).Because we have demonstrated already that cell-collagen V interactions involved integrins (9, 10), the binding of membrane-spanning proteoglycans could reinforce cell attachment to collagen V and, in that sense, would be of physiological importance....

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“…Interdomain interactions are important determinants of the overall tertiary structure of the protein (20 -22). Fibronectin is able to interact with heparin and heparan sulfate, and there is evidence that this interaction affects the structure of the protein (23)(24)(25)(26)(27)(28). In a previous study, we demonstrated that heparin mediates a conformational change in fibronectin from a compact to a more extended conformation, potentially by interfering with interdomain interactions (29).…”

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confidence: 85%

A Catalytic Role of Heparin within the Extracellular Matrix

Mitsi

Forsten-Williams

Gopalakrishnan

et al. 2008

Journal of Biological Chemistry

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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 heparininduced 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.

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Binding of heparin by type III domains and peptides from the carboxy terminal Hep-2 region of fibronectin

Cited by 47 publications

References 26 publications

Mapping the heparin‐binding domain of boar spermadhesins

Mapping the heparin‐binding domain of boar spermadhesins

Molecular Features of the Collagen V Heparin Binding Site

A Catalytic Role of Heparin within the Extracellular Matrix

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