Nanomechanics on FGF-2 and Heparin Reveal Slip Bond Characteristics with pH Dependency

Sevim, Semih; Ozer, Sevil; Jones, Gabriel Davis; Wurzel, Joel; Feng, Luying; Fakhraee, Arielle; Shamsudhin, Naveen; Ergeneman, Olgaç; Pellicer, Eva; Sort, Jordi; Nelson, Bradley J.; Torun, Hamdi; Lühmann, Tessa

doi:10.1021/acsbiomaterials.6b00723

Cited by 7 publications

(9 citation statements)

References 33 publications

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“…The lifetime values are consistent with the responses in neurons (e.g., neurite retraction) triggered by the astrocyte proteins: α V β 3 integrin induces a significant retraction of neuronal processes, while the proteoglycan Syndecan-4 does not promote any morphological changes ( Figure 2D ). The slip bond behavior found for both bi-molecular interactions coincides with the mechanical characterization described for Thy-1–α 5 β 1 binding ( Fiore et al, 2014 ) as well as for FGF with the HSPG ( Sevim et al, 2017 ). Moreover, we found that the ternary complex (Thy-1–α V β 3 integrin + Syndecan-4) is more stable in terms of force-dependent lifetime than each binary complex.…”

Section: Discussionsupporting

confidence: 79%

Application of Force to a Syndecan-4 Containing Complex With Thy-1–αVβ3 Integrin Accelerates Neurite Retraction

Burgos‐Bravo

Martínez-Meza

Quest

et al. 2020

Front. Mol. Biosci.

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Inflammation contributes to the genesis and progression of chronic diseases, such as cancer and neurodegeneration. Upregulation of integrins in astrocytes during inflammation induces neurite retraction by binding to the neuronal protein Thy-1, also known as CD90. Additionally, Thy-1 alters astrocyte contractility and movement by binding to the mechano-sensors α V β 3 integrin and Syndecan-4. However, the contribution of Syndecan-4 to neurite shortening following Thy-1–α V β 3 integrin interaction remains unknown. To further characterize the contribution of Syndecan-4 in Thy-1-dependent neurite outgrowth inhibition and neurite retraction, cell-based assays under pro-inflammatory conditions were performed. In addition, using Optical Tweezers, we studied single-molecule binding properties between these proteins, and their mechanical responses. Syndecan-4 increased the lifetime of Thy-1–α V β 3 integrin binding by interacting directly with Thy-1 and forming a ternary complex (Thy-1–α V β 3 integrin + Syndecan-4). Under in vitro -generated pro-inflammatory conditions, Syndecan-4 accelerated the effect of integrin–engaged Thy-1 by forming this ternary complex, leading to faster neurite retraction and the inhibition of neurite outgrowth. Thus, Syndecan-4 controls neurite cytoskeleton contractility by modulating α V β 3 integrin mechano-receptor function. These results suggest that mechano-transduction, cell-matrix and cell-cell interactions are likely critical events in inflammation-related disease development.

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Section: Discussionsupporting

confidence: 79%

Application of Force to a Syndecan-4 Containing Complex With Thy-1–αVβ3 Integrin Accelerates Neurite Retraction

Burgos‐Bravo

Martínez-Meza

Quest

et al. 2020

Front. Mol. Biosci.

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“…Specifically, we use atomic force microscopy (AFM) based single-molecule force spectroscopy (SMFS) to quantify the response of individual molecular interactions to tensile forces. This force probe technique is now well established for the analysis of intra- and intermolecular forces (29, 30) and is emerging for the probing of glycosaminoglycan-protein interactions (16, 31, 32). A prerequisite of AFM SMFS measurements is the proper immobilization of the molecules to be probed, in a way that permits the controlled application of the necessary tensile forces.…”

Section: Introductionmentioning

confidence: 99%

Single-Molecule Unbinding Forces between the Polysaccharide Hyaluronan and Its Binding Proteins

Bano

Tammi

Kang

et al. 2018

Biophysical Journal

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The extracellular polysaccharide hyaluronan (HA) is ubiquitous in all vertebrate tissues, where its various functions are encoded in the supramolecular complexes and matrices that it forms with HA-binding proteins (hyaladherins). In tissues, these supramolecular architectures are frequently subjected to mechanical stress, yet how this affects the intermolecular bonding is largely unknown. Here, we used a recently developed single-molecule force spectroscopy platform to analyze and compare the mechanical strength of bonds between HA and a panel of hyaladherins from the Link module superfamily, namely the complex of the proteoglycan aggrecan and cartilage link protein, the proteoglycan versican, the inflammation-associated protein TSG-6, the HA receptor for endocytosis (stabilin-2/HARE), and the HA receptor CD44. We find that the resistance to tensile stress for these hyaladherins correlates with the size of the HA-binding domain. The lowest mean rupture forces are observed for members of the type A subgroup (i.e., with the shortest HA-binding domains; TSG-6 and HARE). In contrast, the mechanical stability of the bond formed by aggrecan in complex with cartilage link protein (two members of the type C subgroup, i.e., with the longest HA-binding domains) and HA is equal or even superior to the high affinity streptavidin⋅biotin bond. Implications for the molecular mechanism of unbinding of HA⋅hyaladherin bonds under force are discussed, which underpin the mechanical properties of HA⋅hyaladherin complexes and HA-rich extracellular matrices.

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“…Increasing ks H by four orders of magnitude only decreased cell surface HSPG concentration by about two times. Finally, it has recently been proposed that FGF2 and heparin demonstrate slip-bond characteristics 21 . We therefore varied FGF2-HSPG k off several orders of magnitude above and below the base value (dotted line, Figure 4E).…”

Section: Resultsmentioning

confidence: 99%

“…Computational studies of the endothelial glycocalyx have suggested that shear stress stretches cell surface HSPG [60][61][62] , which would expose additional FGF2 binding sites and increase FGF2 capture by HSPG as FGF2 dimers in flow. Sevim et al investigated the nanomechanics of the FGF2-HSPG bond itself using atomic force microscopy 21 . Their data suggest that FGF2-HSPG dissociation rate was constant at loading rates of 101 to 106 pN/s but then increased approximately 4500 times at higher loading rates.…”

Section: Discussionmentioning

confidence: 99%

“…Recent studies suggest that shear stress upregulates endothelial cell HSPG expression and regrowth as compared to cells in static culture 19,20 . Furthermore, Sevim et al used atomic force microscopy to demonstrate slip bond characteristics between FGF2 and heparin, introducing the possibility that the FGF2-HSPG bond itself may change under flow conditions 21 . It is therefore important to explore how flow-adapted endothelial cells may differentially bind FGF2, including the mechanisms for this effect.…”

Section: Introductionmentioning

confidence: 99%

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Fibroblast Growth Factor-2 Binding to Heparan Sulfate Proteoglycans Varies with Shear Stress in Flow-Adapted Cells

et al. 2019

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Fibroblast Growth Factor 2 (FGF2), an important regulator of angiogenesis, binds to endothelial cell (EC) surface fibroblast growth factor receptors (FGFR) and heparan sulfate proteoglycans (HSPG). FGF2 binding kinetics have been predominantly studied in static culture; however, the endothelium is constantly exposed to flow which may affect FGF2 binding. We therefore used experimental and computational techniques to study how EC FGF2 binding changes in flow. ECs adapted to 24 hours of flow demonstrated biphasic FGF2-HSPG binding, with FGF2-HSPG complexes increasing up to 20 dynes/cm 2 shear stress and then decreasing at higher shear stresses. To understand how adaptive EC surface remodeling in response to shear stress may affect FGF2 binding to FGFR and HSPG, we implemented a computational model to predict the relative effects of flow-induced surface receptor changes. We then fit the computational model to the experimental data using relationships between HSPG availability and FGF2-HSPG dissociation and flow that were developed from a basement membrane study, as well as including HSPG production. These studies suggest that FGF2 binding kinetics are altered in flow-adapted ECs due to changes in cell surface receptor quantity, availability, and binding kinetics, which may affect cell growth factor response.

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Nanomechanics on FGF-2 and Heparin Reveal Slip Bond Characteristics with pH Dependency

Cited by 7 publications

References 33 publications

Application of Force to a Syndecan-4 Containing Complex With Thy-1–αVβ3 Integrin Accelerates Neurite Retraction

Application of Force to a Syndecan-4 Containing Complex With Thy-1–αVβ3 Integrin Accelerates Neurite Retraction

Single-Molecule Unbinding Forces between the Polysaccharide Hyaluronan and Its Binding Proteins

Fibroblast Growth Factor-2 Binding to Heparan Sulfate Proteoglycans Varies with Shear Stress in Flow-Adapted Cells

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