Dataset Information

Fibronectin binding to von Willebrand factor occurs via the A1 domain.

ABSTRACT:

Background

Collagen interactions with von Willebrand factor (VWF) perform an important role in initiation of hemostasis.

Objectives

We hypothesized that in addition to collagen, other extracellular matrix (ECM) proteins such as fibronectin can bind VWF.

Methods

Fibronectin-VWF interactions were measured by ELISA using both plasma-derived and recombinant VWF-containing variants in specific domains. Inhibition was measured by antibody competition using antibodies directed against both VWF and fibronectin. Binding affinities were measured by the Octet Biosensor for fibronectin and collagen IV.

Results

Fibronectin was able to bind both plasma-derived and recombinant wild-type VWF. This interaction was inhibited by both anti-VWF antibodies and collagen types III and IV. Several VWF A1 domain variants in the region of the collagen IV binding site also demonstrated absent fibronectin binding, as did variants with defects in high-molecular-weight multimers. Binding affinity testing showed fibronectin has a strong affinity for VWF, in a range similar to that of collagen IV. Fibronectin binds VWF via a restricted region of the A1 domain. This interaction requires high-molecular-weight multimers and is similar to that seen with vascular collagens.

Conclusions

Therefore, VWF would appear to be the common factor linking platelet adhesion to various ECM proteins and facilitating hemostasis under conditions of ECM exposure.

SUBMITTER: Keesler DA

PROVIDER: S-EPMC8178691 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Publications

Fibronectin binding to von Willebrand factor occurs via the A1 domain.

Keesler Daniel A DA Slobodianuk Tricia L TL Kochelek Caroline E CE Skaer Chad W CW Haberichter Sandra L SL Flood Veronica H VH

Research and practice in thrombosis and haemostasis 20210605 5

<h4>Background</h4>Collagen interactions with von Willebrand factor (VWF) perform an important role in initiation of hemostasis.<h4>Objectives</h4>We hypothesized that in addition to collagen, other extracellular matrix (ECM) proteins such as fibronectin can bind VWF.<h4>Methods</h4>Fibronectin-VWF interactions were measured by ELISA using both plasma-derived and recombinant VWF-containing variants in specific domains. Inhibition was measured by antibody competition using antibodies directed aga ...[more]

PMID: 34136746

Similar Datasets

Project description:Essentials The mechanism for the auto-inhibition of von Willebrand factor (VWF) remains unclear. Hydrogen exchange of two VWF A1 fragments with disparate activities was measured and compared. Discontinuous residues flanking A1 form a structural module that blocks A1 binding to the platelet. Our results suggest a potentially unified model of VWF activation. Click to hear an ISTH Academy presentation on the domain architecture of VWF and activation by elongational flow by Dr Springer SUMMARY: Background How von Willebrand factor (VWF) senses and responds to shear flow remains unclear. In the absence of shear flow, VWF or its fragments can be induced to bind spontaneously to platelet GPIbα. Objectives To elucidate the auto-inhibition mechanism of VWF. Methods Hydrogen-deuterium exchange (HDX) of two recombinant VWF fragments expressed from baby hamster kidney cells were measured and compared. Results The shortA1 protein contains VWF residues 1261-1472 and binds GPIbα with a significantly higher affinity than the longA1 protein that contains VWF residues 1238-1472. Both proteins contain the VWF A1 domain (residues 1272-1458). Many residues in longA1, particularly those in the N- and C-terminal sequences flanking the A1 domain, and in helix α1, loops α1β2 and β3α2, demonstrated markedly reduced HDX compared with their counterparts in shortA1. The HDX-protected region in longA1 overlaps with the GPIbα-binding interface and is clustered with type 2B von Willebrand disease (VWD) mutations. Additional comparison with the HDX of denatured longA1 and ristocetin-bound longA1 indicates the N- and C-terminal sequences flanking the A1 domain form cooperatively an integrated autoinhibitory module (AIM) that interacts with the HDX-protected region. Binding of ristocetin to the C-terminal part of the AIM desorbs the AIM from A1 and enables longA1 binding to GPIbα. Conclusion The discontinuous AIM binds the A1 domain and prevents it from binding to GPIbα, which has significant implications for the pathogenesis of type 2B VWD and the shear-induced activation of VWF activity.

Dataset Information

Fibronectin binding to von Willebrand factor occurs via the A1 domain.

Background

Objectives

Methods

Results

Conclusions

Publications

Fibronectin binding to von Willebrand factor occurs via the A1 domain.

Similar Datasets

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