Single-molecule dynamic force spectroscopy of the fibronectin–heparin interaction

Mitchell, Gabriel; Lamontagne, Charles-Antoine; Lebel, Réjean; Grandbois, Michel; Malouin, François

doi:10.1016/j.bbrc.2007.10.034

Cited by 10 publications

(14 citation statements)

References 29 publications

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“…Human plasma fibronectin (Calbiochem, ON, Canada) was covalently attached to silicon nitride tips (Veeco, CA, USA) using carboxymethylamylose (CMA) spacers (Sigma‐Aldrich) and the carbodiimide chemistry as previously described (Mitchell et al ., 2007). The CMA was used as a molecular spacer because non‐specific interactions between the bacteria and the CMA‐modified probe without fibronectin were rare and did not have the same properties as those of the studied interaction.…”

Section: Methodssupporting

confidence: 75%

“…The resistance and the compliance of a particular molecular interaction to a mechanical load can thus be measured by varying the applied force. Since there is a correlation between the loading rate of an applied force and the mean unbinding force of a particular molecular interaction, it is possible to study the mechanical response of an interaction to various external forces by doing experiments at different loading rates (see Experimental procedures ) (Evans and Ritchie, 1997; Mitchell et al ., 2007). Figure 3 presents the dynamic force spectrum or the mean unbinding force as a function of the logarithm of the mean loading rate of the fibronectin–FnBPs interaction obtained with the Newbould strain in early exponential growth phase.…”

Section: Resultsmentioning

confidence: 99%

“…Briefly, there is usually a correlation between the logarithm of the loading rate and the mean unbinding force of a particular molecular interaction. This is described by the following equation (Evans and Ritchie, 1997; Mitchell et al ., 2007):…”

Section: Methodsmentioning

confidence: 99%

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Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Mitchell

Lamontagne

Brouillette

et al. 2008

Molecular Microbiology

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SummaryGenes encoding cell-surface proteins regulated by SigB are stably expressed in Staphylococcus aureus small-colony variants (SCVs) isolated from cystic fibrosis (CF) patients. Our hypothesis is that CF-isolated SCVs are locked into a colonization state by sustaining the expression of adhesins such as fibronectin-binding proteins (FnBPs) throughout growth. Force spectroscopy was used to study the fibronectin-FnBPs interaction among strains varying for their SigB activity. The fibronectin-FnBPs interaction was described by a strength of 1000 Ϯ 400 pN (pulling rate of 2 mm s . A CF-isolated SCV highly expressed fnbA throughout growth and showed a sustained capacity to bind fibronectin, whereas a prototypic strain showed a reduced frequency of fibronectin-binding during the stationary growth phase when its fnbA gene was down-regulated. Reduced expression of fnbA was observed in sigB mutants, which was associated with an overall decrease adhesion to fibronectin. These results suggest that the fibronectin-FnBPs interaction plays a role in the formation of a mechanically resistant adhesion of S. aureus to host tissues and supports the hypothesis that CF-isolated SCVs are locked into a colonization state as a result of a sustained SigB activity.

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Section: Methodssupporting

confidence: 75%

Section: Resultsmentioning

confidence: 99%

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Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Mitchell

Lamontagne

Brouillette

et al. 2008

Molecular Microbiology

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“…45,49 Barkalow and Schwarzbauer 50 and others have previously reported the binding of heparin to fibronectin and with other molecules of extracellular matrix. 10,[51][52][53][54][55] Based on this literature, we hypothesized that heparin could interact indirectly with M 3 receptor through integrin. Our results showed that heparin induced activation of focal adhesion proteins involved in integrin signaling ( Figure 5A through 5C).…”

Section: Discussionmentioning

confidence: 99%

Heparin Induces Rat Aorta Relaxation via Integrin-Dependent Activation of Muscarinic M ₃ Receptors

et al. 2010

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Abstract-Previous reports have shown that heparin may promote human hypotension and vascular relaxation by elevation of NO levels through unclear mechanisms. We hypothesized that endothelial muscarinic M 3 receptor activation mediates the heparin-induced vasodilation of rat aortic rings. The experiments were carried out using unfractionated heparin extracted from bovine intestinal mucosa, which elicited an endothelium and NO-dependent relaxation of aortic segments with maximal potency and efficacy (EC 50 : 100Ϯ10 mol/L; E max : 41Ϯ3%). Atropine and 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide inhibitors reduced the heparin-dependent relaxation, indicating that M 3 muscarinic receptor is involved in this phenomenon. However, no direct binding of heparin to muscarinic receptors was observed.More importantly, studies performed using the arginine-glycine-aspartic acid peptide and 1-(1,1-dimethylethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-day]pyrimidin-4-amine, an Src family inhibitor, reduced by 51% and 73% the heparin-dependent relaxation, respectively, suggesting the coupling of heparin and M 3 receptor through extracellular matrix molecules and integrin. Furthermore, unfractionated heparin induced activation of focal adhesion protein kinase, Src, and paxillin. Finally, fluorescence resonance energy transfer approach confirmed the interaction of the M 3 receptor to integrin. Taken together, these data demonstrate the participation of M 3 receptor and integrin in heparin-dependent relaxation of vascular smooth muscle. These results provide new insights into the molecular mechanism and potential pharmacological action of heparin in vascular physiology. (Hypertension. 2010;56:713-721.)Key Words: heparin Ⅲ muscarinic receptors Ⅲ M 3 receptor Ⅲ integrin Ⅲ smooth muscle relaxation Ⅲ aorta T he successful use of heparin in the treatment of thromboembolic diseases has been associated with numerous pharmacological actions of this complex polysaccharide. Alterations in the blood vessel and associated cells, such as platelet hyperactivity, inflammatory processes, endothelial dysfunction, and angiogenesis, lead to the development of diseases related to clot formation, which can be efficiently modulated by heparin and its derivatives. [1][2][3] Heparin is a glycosaminoglycan composed of repeating 13 4-linked ␣-D-glucosamine, mainly N-sulfated, and uronic acid, either ␤-D-glucuronic acid or ␣-L-iduronic acid; also, O-sulfation occurs at different positions of the disaccharide units. 4,5 Although heparin usually acts as an anticoagulant, 6 other biological activities have also been described, including antiviral, 7 antibacterial, 8 and antithrombotic effects. This antithrombotic action has been related, at least in part, to the increased production of a peculiar heparan sulfate proteoglycan produced by the endothelium. 9 -11 In addition, heparin modulates the function of many proteins, for example, the myosin from skeletal muscle, 12 the inositol 1,4,5-trisphosphate receptor inhibition, 13 and the activation of Na ϩ /Ca 2ϩ ...

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“…1. Force spectroscopy experiments have previously been used to characterize binding between fibronectin and heparin (32), and these are compared with the current results. It is shown that the energy barrier but not the dissociation rate constant is similar for SDC4 and heparin.…”

Section: Introductionmentioning

confidence: 99%

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

Kennelly

Qwarnström

et al. 2019

Biophysical Journal

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Dynamic single-molecule force spectroscopy was performed to monitor the unbinding of fibronectin with the proteoglycans syndecan-4 (SDC4) and decorin and to compare this with the unbinding characteristics of a 5 b 1-integrin. A single energy barrier was sufficient to describe the unbinding of both SDC4 and decorin from fibronectin, whereas two barriers were observed for the dissociation of a 5 b 1-integrin from fibronectin. The outer (high-affinity) barriers in the interactions of fibronectin with a 5 b 1-integrin and SDC4 are characterized by larger barrier heights and widths and slower dissociation rates than those of the inner (low-affinity) barriers in the interactions of fibronectin with a 5 b 1-integrin and decorin. These results indicate that SDC4 and (ultimately) a 5 b 1-integrin have the ability to withstand deformation in their interactions with fibronectin, whereas the decorinfibronectin interaction is considerably more brittle.

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Single-molecule dynamic force spectroscopy of the fibronectin–heparin interaction

Cited by 10 publications

References 29 publications

Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Heparin Induces Rat Aorta Relaxation via Integrin-Dependent Activation of Muscarinic M ₃ Receptors

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

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Single-molecule dynamic force spectroscopy of the fibronectin–heparin interaction

Cited by 10 publications

References 29 publications

Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Staphylococcus aureus SigB activity promotes a strong fibronectin–bacterium interaction which may sustain host tissue colonization by small‐colony variants isolated from cystic fibrosis patients

Heparin Induces Rat Aorta Relaxation via Integrin-Dependent Activation of Muscarinic M 3 Receptors

Distinct Binding Interactions of α5β1-Integrin and Proteoglycans with Fibronectin

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Heparin Induces Rat Aorta Relaxation via Integrin-Dependent Activation of Muscarinic M ₃ Receptors