Cooperation within von Willebrand factors enhances adsorption mechanism

von Willebrand factor (VWF) is a naturally collapsed protein that participates in primary haemostasis and coagulation events. The clotting process is triggered by the adsorption and conformational changes of the plasma VWFs localized to the collagen fibres found near the site of injury. We develop coarse-grained models to simulate the adsorption dynamics of VWF flowing near the adhesive collagen fibres at different shear rates and investigate the effect of factors such as interaction and cooperativity of VWFs on the success of adsorption events. The adsorption probability of a flowing VWF confined to the receptor field is enhanced when it encounters an adhered VWF in proximity to the collagen receptors. This enhancement is observed within a wide range of shear rates and is mostly controlled by the attractive van der Waals interactions rather than the hydrodynamic interactions among VWF monomers. The cooperativity between the VWFs acts as an effective mechanism for enhancing VWF adsorption to the collagen fibres. Additionally, this implies that the adsorption of such molecules is nonlinearly dependent on the density of flowing VWFs. These findings are important for studies of primary haemostasis as well as general adsorption dynamics processes in polymer physics.

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Platelet function under high shear conditions

Hämostaseologie ◽

10.1055/s-0037-1616934 ◽

2009 ◽

Vol 29 (01) ◽

pp. 21-24 ◽

Cited By ~ 9

Author(s):

A. J. Reininger

Keyword(s):

Blood Platelets ◽

Immunoglobulin Superfamily ◽

High Shear ◽

Shear Rates ◽

Collagen Receptors ◽

High Shear Rates ◽

Platelet Receptor ◽

Flowing Blood ◽

Von Willebrand ◽

Willebrand Factor

SummaryBlood platelets are the first line of defense against bleeding and as such involved in the haemostatic repair of damaged vasculature. Their true prowess seems to be displayed under high shear conditions where platelets interact with a variety of plasma proteins, all of which are tightly regulated to close the leak but at the same time prevent lumen occlusion and thromboembolism. The first task is to arrest fast flowing platelets on exposed collagen of the damaged subendothelial surface. Although platelets are endowed with several collagen receptors, most notably integrin ╒2b®1 and the immunoglobulin superfamily member GPVI, they can not arrest platelets at high shear rates. The latter requires binding of the platelet receptor GPIb╒to the A1-binding domain of von Willebrand factor (VWF), which first has to be immobilized from the flowing blood onto the site of injury. Under high shear conditions further accrual of newly arriving platelets again requires VWF, which has to bridge platelets not only to the exposed collagen but also to each other by being sandwiched between the multiple platelet layers of the haemostatic plug.

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Distinct Functional Conformations of von Willebrand Factor A1 Domain in Support of Platelet-Surface or Platelet-Platelet Interactions.

Blood ◽

10.1182/blood.v110.11.5182.5182 ◽

2007 ◽

Vol 110 (11) ◽

pp. 5182-5182

Author(s):

Gianmarco Podda ◽

James R. Roberts ◽

Richard A. McClintock ◽

Zaverio M. Ruggeri

Keyword(s):

Platelet Aggregation ◽

Von Willebrand Factor ◽

Conformational Changes ◽

Platelet Adhesion ◽

Shear Rates ◽

Amino Terminal ◽

A1 Domain ◽

Von Willebrand ◽

Willebrand Factor ◽

Positively Charged

Abstract The adhesive protein, von Willebrand factor (VWF), is generally considered a key substrate for platelet adhesion to the vessel wall, yet its role in platelet cohesion (aggregation) may be equally important for normal thrombus formation. In either case, the function of VWF is mediated by the primary interaction of the VWF A1 domain (VWF-A1) with glycoprotein (GP) Ibα, a component of the GPIb-IX-V receptor complex on the platelet membrane. Because normal plasma VWF in solution and GPIb coexist in circulating blood without any appreciable interaction, it has been postulated that conformational changes occur when VWF becomes immobilized and/or under the effect of pathologically elevated shear stress, such that binding to the receptor becomes possible and resultis in platelet tethering to a surface and shear-induced aggregation. Changes of the molecular shape of VWF, from coiled to extended, have been shown under the effect of hemodynamic forces, but evidence for conformational changes within VWF-A1 has remained elusive. The crystal structure of VWF-A1 in complex with a GPIbα amino terminal fragment has revealed that the VWF-A1 residues involved in the interaction are comprised between positions 544–614 and, in particular, do not include several positively charged Arg and Lys residues located in helices α4 and 5 (residues 627–668). The latter appear as likely candidates to interact with negatively charged residues in GPIbα as a consequence of potential conformational changes induced by tensile stress on the bond following an initial ligand-receptor contact. We tested this hypothesis by evaluating the ability of selected VWF-A1 mutants to support platelet adhesion or aggregation, respectively, under controlled flow conditions. Methods. We expressed in insect cells and purified a series of VWF-A1 fragments comprising residues 445–733. One fragment had native sequence and 8 had single or multiple substitutions of positively charged amino acid residues in helices α4 and/or α5. None of the substituted residues contribute to contacts with GP Ibα in the known crystal structures of the corresponding complex, and all except one were between 8 and 20 angstroms away from the closest GPIbα residue. All the fragments were dimeric (d) owing to the presence of interchain disulfide bond(s). Results: Native dVWF-A1 in solution supported platelet aggregation in a laminar flow field. Of the 8 mutants, 5 had variably decreased function (up to 95% less aggregation) and 2 had increased function (up to 200% increase in aggregation). The same results were observed with platelet-rich plasma in suspension or by measuring platelet aggregate formation with blood cells perfused over immobilized VWF-A1 at wall shear rates as high as 10,000 1/s. In contrast, as judged by the number of tethered platelets and their rolling velocities, all mutants supported adhesion as well as or better that the native VWFA-1 at all shear rates tested (500–25,000 1/s). Conclusions: These results provide structural evidence for the existence of different VWF-A1 conformers that can modulate adhesive properties with distinct effects on platelet adhesion to a surface or platelet aggregation.

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Synergism between platelet collagen receptors defined using receptor-specific collagen-mimetic peptide substrata in flowing blood

Blood ◽

10.1182/blood-2010-01-260778 ◽

2010 ◽

Vol 115 (24) ◽

pp. 5069-5079 ◽

Cited By ~ 79

Author(s):

Nicholas Pugh ◽

Anna M. C. Simpson ◽

Peter A. Smethurst ◽

Philip G. de Groot ◽

Nicolas Raynal ◽

...

Keyword(s):

Thrombus Formation ◽

Confocal Imaging ◽

Peptide Substrates ◽

Shear Rates ◽

Glycoprotein Vi ◽

Collagen Receptors ◽

Platelet Binding ◽

Von Willebrand ◽

Willebrand Factor

AbstractExposed subendothelial collagen acts as a substrate for platelet adhesion and thrombus formation after vascular injury. Synthetic collagen-derived triple-helical peptides, designated collagen-related peptide (CRP), GFOGER, and VWF-III, can specifically engage the platelet collagen receptors, glycoprotein VI and integrin α2β1, and plasma von Willebrand factor (VWF), respectively. Hitherto, the role of these 3 collagen-binding axes has been studied indirectly. Use of these uniform peptide substrates, rather than collagen fibers, provides independent control of each axis. Here, we use confocal imaging and novel image analysis techniques to investigate the effects of receptor-ligand engagement on platelet binding and activation during thrombus formation under flow conditions. At low shear (100s−1 and 300s−1), both GFOGER and CRP are required for thrombus formation. At 1000s−1, a combination of either CRP or GFOGER with VWF-III induces comparable thrombus formation, and VWF-III increases thrombus deposition at all shear rates, being indispensable at 3000s−1. A combination of CRP and VWF-III is sufficient to support extensive platelet deposition at 3000s−1, with slight additional effect of GFOGER. Measurement of thrombus height after specific receptor blockade or use of altered proportions of peptides indicates a signaling rather than adhesive role for glycoprotein VI, and primarily adhesive roles for both α2β1 and the VWF axis.

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Predictive relation for the α-relaxation time of a coarse-grained polymer melt under steady shear

Science Advances ◽

10.1126/sciadv.aaz0777 ◽

2020 ◽

Vol 6 (17) ◽

pp. eaaz0777 ◽

Cited By ~ 3

Author(s):

Andrea Giuntoli ◽

Francesco Puosi ◽

Dino Leporini ◽

Francis W. Starr ◽

Jack F. Douglas

Keyword(s):

Relaxation Time ◽

Structural Relaxation ◽

Polymer Melt ◽

Steady Shear ◽

Coarse Grained ◽

Scattering Function ◽

Dynamic Heterogeneity ◽

Shear Rates ◽

Wide Range ◽

Intermediate Scattering Function

We examine the influence of steady shear on structural relaxation in a simulated coarse-grained unentangled polymer melt over a wide range of temperature and shear rates. Shear is found to progressively suppress the α-relaxation process observed in the intermediate scattering function, leading ultimately to a purely inertially dominated β-relaxation at high shear rates, a trend similar to increasing temperature. On the basis of a scaling argument emphasizing dynamic heterogeneity in cooled liquids and its alteration under material deformation, we deduce and validate a parameter-free scaling relation for both the structural relaxation time τα from the intermediate scattering function and the “stretching exponent” β quantifying the extent of dynamic heterogeneity over the entire range of temperatures and shear rates that we can simulate.

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Electrostatics and Solvation: Essential Determinants of Chromatin Compaction

10.1101/785634 ◽

2019 ◽

Author(s):

A. Bendandi ◽

S. Dante ◽

S. Rehana Zia ◽

A. Diaspro ◽

W. Rocchia

Keyword(s):

Conformational Changes ◽

Protein Interactions ◽

Electrostatic Interactions ◽

Ionic Concentration ◽

Coarse Grained ◽

Chromatin Compaction ◽

Spatial And Temporal Scales ◽

Chromatin Fibre ◽

Nucleosome Core ◽

Wide Range

ABSTRACTChromatin compaction is a process of fundamental importance in Biology, as it greatly influences cellular function and gene expression. The dynamics of compaction is determined by the interactions between DNA and histones, which are mainly mechanical and electrostatic. The high charge of DNA makes electrostatics extremely important for chromatin topology and dynamics. Besides their mechanical and steric role in the chromatin fibre, linker DNA length and linker histone presence and binding position also bear great electrostatic consequences. Electrostatics in chromatin is also indirectly linked to the DNA sequence: the presence of high-curvature AT-rich segments in DNA can cause conformational variations with electrostatic repercussions, attesting to the fact that the role of DNA is both structural and electrostatic. Electrostatics in this system has been analysed by extensively examining at the computational level the repercussions of varying ionic concentration, using all-atom, coarse-grained, and continuum models. There have been some tentative attempts to describe the force fields governing chromatin conformational changes and the energy landscapes of these transitions, but the intricacy of the system has hampered reaching a consensus. Chromatin compaction is a very complex issue, depending on many factors and spanning orders of magnitude in space and time in its dynamics. Therefore, comparison and complementation of theoretical models with experimental results is fundamental. Here, we present existing approaches to analyse electrostatics in chromatin and the different points of view from which this issue is treated. We pay particular attention to solvation, often overlooked in chromatin studies. We also present some numerical results on the solvation of nucleosome core particles. We discuss experimental techniques that have been combined with computational approaches and present some related experimental data such as the Z-potential of nucleosomes at varying ionic concentrations. Finally, we discuss how these observations support the importance of electrostatics and solvation in chromatin models.SIGNIFICANCEThis work explores the determinants of chromatin compaction, focusing on the importance of electrostatic interactions and solvation. Chromatin compaction is an intrinsically multiscale issue, since processes concerning chromatin occur on a wide range of spatial and temporal scales. Since DNA is a highly charged macromolecule, electrostatic interactions are extremely significant for chromatin compaction, an effect examined in this work from many angles, such as the importance of ionic concentration and different ionic types, DNA-protein interactions, and solvation. Solvation is often overlooked in chromatin studies, especially in coarse-grained models, where the nucleosome core, the building block of the chromatin fibre, is represented as a rigid body, even though it has been observed that solvation influences chromatin even at the base-pair level.

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Emergence of ion channel modal gating from independent subunit kinetics

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1604090113 ◽

2016 ◽

Vol 113 (36) ◽

pp. E5288-E5297 ◽

Cited By ~ 12

Author(s):

Brendan A. Bicknell ◽

Geoffrey J. Goodhill

Keyword(s):

Ion Channels ◽

Conformational Changes ◽

Ionic Currents ◽

Analytical Framework ◽

Stochastic Simulations ◽

Coarse Grained ◽

Wide Range ◽

Intracellular Ca ◽

Modal Gating ◽

Kinetics Of

Many ion channels exhibit a slow stochastic switching between distinct modes of gating activity. This feature of channel behavior has pronounced implications for the dynamics of ionic currents and the signaling pathways that they regulate. A canonical example is the inositol 1,4,5-trisphosphate receptor (IP3R) channel, whose regulation of intracellular Ca2+ concentration is essential for numerous cellular processes. However, the underlying biophysical mechanisms that give rise to modal gating in this and most other channels remain unknown. Although ion channels are composed of protein subunits, previous mathematical models of modal gating are coarse grained at the level of whole-channel states, limiting further dialogue between theory and experiment. Here we propose an origin for modal gating, by modeling the kinetics of ligand binding and conformational change in the IP3R at the subunit level. We find good agreement with experimental data over a wide range of ligand concentrations, accounting for equilibrium channel properties, transient responses to changing ligand conditions, and modal gating statistics. We show how this can be understood within a simple analytical framework and confirm our results with stochastic simulations. The model assumes that channel subunits are independent, demonstrating that cooperative binding or concerted conformational changes are not required for modal gating. Moreover, the model embodies a generally applicable principle: If a timescale separation exists in the kinetics of individual subunits, then modal gating can arise as an emergent property of channel behavior.

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Platelet Collagen Receptors

Thrombosis and Haemostasis ◽

10.1055/s-0037-1616217 ◽

2001 ◽

Vol 86 (07) ◽

pp. 189-197 ◽

Cited By ~ 182

Author(s):

Jeannine Clemetson ◽

Kenneth Clemetson

Keyword(s):

Platelet Activation ◽

Collagen Types ◽

Shear Rates ◽

Collagen Receptors ◽

High Shear Rates ◽

Ig Superfamily ◽

Von Willebrand ◽

Willebrand Factor ◽

Collagen Receptor ◽

Integrin Α2

SummaryCollagens are important platelet activators in the vascular subendothelium and vessel wall. Since the regulation of platelet activation is a key step in distinguishing normal haemostasis from pathological thrombosis, collagen interactions with platelets are important targets for pharmacological control. Platelets have two major receptors for collagens, the integrin α2 β1, with a major role in adhesion and platelet anchoring and the Ig superfamily member, GPVI, principally responsible for signalling and platelet activation. In addition, GPIb-V-IX, can be considered as an indirect collagen receptor acting via von Willebrand factor as bridging molecule and is essential for platelet interactions with collagen at high shear rates. There is some evidence for additional receptors, which may regulate the response to individual collagen types. This review discusses how these receptors work separately with specific agonists and proposes possible mechanisms for how they work together to regulate platelet activation by collagen, which remains controversial and poorly understood.

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Asialo von Willebrand Factor Enhances Platelet Adhesion to Vessel Subendothelium

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647629 ◽

1988 ◽

Vol 60 (01) ◽

pp. 030-034 ◽

Cited By ~ 9

Author(s):

Eva Bastida ◽

Juan Monteagudo ◽

Antonio Ordinas ◽

Luigi De Marco ◽

Ricardo Castillo

Keyword(s):

Von Willebrand Factor ◽

Platelet Adhesion ◽

Human Albumin ◽

Membrane Receptors ◽

Shear Rates ◽

High Shear Rates ◽

Platelet Deposition ◽

Platelet Spreading ◽

Von Willebrand ◽

Willebrand Factor

SummaryNative von Willebrand factor (N-vWF) binds to platelets activated by thrombin, ADP or ristocetin. Asialo vWF (As-vWF) induces platelet aggregation in absence of platelet activators. N-vWF mediates platelet adhesion to vessel subendothelium at high shear rates. We have investigated the role of As-vWF in supporting platelet deposition to rabbit vessel subendothelium at a shear rate of 2,000 sec-1, using the Baumgartner perfusion system. We have studied the effects of the addition of As-vWF (from 2 to 12 μg/ml) to perfusates consisting of washed red blood cells, 4% human albumin and washed platelets. Our results show a significant increase in platelet deposition on subendothelium (p <0.01) in perfusions to which As-vWF had been added. Blockage of the platelet glycoproteins Ib and IIb/IIIa (GPIb and GPIIb/IIIa) by specific monoclonal antibodies (LJIb1 and LJCP8, respectively) resulted in a decrease of platelet deposition in both types of perfusates prepared with N-vWF and As-vWF. Our results indicate that As-vWF enhances platelet deposition to vessel subendothelium under flow conditions. Furthermore, they suggest that this effect is mediated by the binding of As-vWF to platelet membrane receptors, which in turn, promote platelet spreading and adhesion to the subendothelium.

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Interventional Thermal Injury of the Arterial Wall: Unfolding of von Willebrand Factor and Its Increased Binding to Collagen after 55° C Heating

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650307 ◽

1996 ◽

Vol 75 (03) ◽

pp. 515-519 ◽

Cited By ~ 7

Author(s):

Mark J Post ◽

Anke N de Graaf-Bos ◽

George Posthuma ◽

Philip G de Groot ◽

Jan J Sixma ◽

...

Keyword(s):

Von Willebrand Factor ◽

Conformational Changes ◽

Arterial Wall ◽

Platelet Adhesion ◽

Type I ◽

Temperature Dependent ◽

Collagen Types ◽

Electron Microscopic ◽

Von Willebrand ◽

Willebrand Factor

Summary Purpose. Thermal angioplasty alters the thrombogenicity of the arterial wall. In previous studies, platelet adhesion was found to increase after heating human subendothelium to 55° C and decrease after heating to 90° C. In the present electron microscopic study, the mechanism of this temperature-dependent platelet adhesion to the heated arterial wall is elucidated by investigating temperature-dependent conformational changes of von Willebrand factor (vWF) and collagen types I and III and the binding of vWF to heated collagen. Methods. Purified vWF and/or collagen was applied to electron microscopic grids and heated by floating on a salt-solution of 37° C, 55° C or 90° C for 15 s. After incubation with a polyclonal antibody against vWF and incubation with protein A/gold, the grids were examined by electron microscopy. Results. At 37° C, vWF was coiled. At 55° C, vWF unfolded, whereas heating at 90° C caused a reduction in antigenicity. Collagen fibers heated to 37° C were 60.3 ± 3.1 nm wide. Heating to 55° C resulted in the unwinding of the fibers, increasing the width to 87.5 ± 8.2 nm (p < 0.01). Heating to 90° C resulted in denatured fibers with an enlarged width of 85.1 ± 6.1 nm (p < 0.05). Heating of collagen to 55° C resulted in an increased vWF binding as compared to collagen heated to 37° C or to 90° C. Incubation of collagen with vWF, prior to heating, resulted in a vWF binding after heating to 55° C that was similar to the 37° C binding and a decreased binding after 90° C. Conclusions. After 55° C heating, the von Willebrand factor molecule unfolds and collagen types I and III exhibit an increased adhesiveness for von Willebrand factor. Heating to 90° C denatures von Willebrand factor and collagen. The conformation changes of von Willebrand factor and its altered binding to collagen type I and III may explain the increased and decreased platelet adhesion to subendothelium after 55° C and 90° C heating, respectively.

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Patterns in diversity and composition of the microbenthos of subarctic intertidal beaches with different morphodynamics

Marine Ecology Progress Series ◽

10.3354/meps13419 ◽

2020 ◽

Vol 648 ◽

pp. 19-38

Author(s):

AI Azovsky ◽

YA Mazei ◽

MA Saburova ◽

PV Sapozhnikov

Keyword(s):

Species Abundance ◽

Abundance Ratio ◽

Wave Action ◽

Coarse Grained ◽

Sediment Properties ◽

Β Diversity ◽

Α Diversity ◽

Wide Range ◽

Similar Mode ◽

Abundance And Diversity

Diversity and composition of benthic diatom algae and ciliates were studied at several beaches along the White and Barents seas: from highly exposed, reflective beaches with coarse-grained sands to sheltered, dissipative silty-sandy flats. For diatoms, the epipelic to epipsammic species abundance ratio was significantly correlated with the beach index and mean particle size, while neither α-diversity measures nor mean cell length were related to beach properties. In contrast, most of the characteristics of ciliate assemblages (diversity, total abundance and biomass, mean individual weight and percentage of karyorelictids) demonstrated a strong correlation to beach properties, remaining low at exposed beaches but increasing sharply in more sheltered conditions. β-diversity did not correlate with beach properties for either diatoms or ciliates. We suggest that wave action and sediment properties are the main drivers controlling the diversity and composition of the intertidal microbenthos. Diatoms and ciliates, however, demonstrated divergent response to these factors. Epipelic and epipsammic diatoms exhibited 2 different strategies to adapt to their environments and therefore were complementarily distributed along the environmental gradient and compensated for each other in diversity. Most ciliates demonstrated a similar mode of habitat selection but differed in their degree of tolerance. Euryporal (including mesoporal) species were relatively tolerant to wave action and therefore occurred under a wide range of beach conditions, though their abundance and diversity were highest in fine, relatively stable sediments on sheltered beaches, whereas the specific interstitial (i.e. genuine microporal) species were mostly restricted to only these habitats.

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