Activation of von Willebrand factor via mechanical unfolding of its discontinuous autoinhibitory module

Arce, Nicholas A.; Cao, Wenpeng; Brown, Alexander K.; Legan, Emily R.; Wilson, Moriah S.; Xu, Emma-Ruoqi; Berndt, Michael C.; Emsley, Jonas; Zhang, Xiao Hui; Li, Renhao

doi:10.1038/s41467-021-22634-x

Cited by 38 publications

(91 citation statements)

References 88 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover, caplacizumab, a bivalent nanobody targeting VWF, was approved by the FDA in 2019 for treating acquired TTP (aTTP) ( 44 , 45 ), a TTP deficiency due to the presence of inhibitory autoantibodies. A recent study indicated that caplacizumab binds the A1 domain (i.e., the platelet-binding domain) of VWF and allosterically inhibits the A1-platelet GPIbα receptor interaction under low and moderate shear, thereby exerting its antithrombotic function ( 46 ).…”

Section: Hemostatic Role Of Vwf/adamts13 Axismentioning

confidence: 99%

Contribution of the von Willebrand factor/ADAMTS13 imbalance to COVID-19 coagulopathy

Seth

McKinnon

Zhang

2022

American Journal of Physiology-Heart and Circulatory Physiology

Self Cite

View full text Add to dashboard Cite

The 2019 coronavirus disease (COVID-19) is the disease caused by SARS-CoV-2 infection. While this infection has been shown to affect the respiratory system, a high incidence of thrombotic events has been observed in severe cases of COVID-19 and in a significant portion of COVID-19 non-survivors. While prior literature has reported on both the coagulopathy and hypercoagulability of COVID-19, the specifics of coagulation have not been fully investigated. Observations of microthrombosis in COVID-19 patients have brought attention to potential inflammatory endothelial injury. Von Willebrand factor (VWF) and its protease, a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13), play an important homeostatic role in responding to endothelial injury. This report provides an overview of the literature investigating the role the VWF/ADAMTS13 axis may have in COVID-19 thrombotic events and suggests potential therapeutic strategies to prevent the progression of coagulopathy in COVID-19 patients.

show abstract

Section: Hemostatic Role Of Vwf/adamts13 Axismentioning

confidence: 99%

Contribution of the von Willebrand factor/ADAMTS13 imbalance to COVID-19 coagulopathy

Seth

McKinnon

Zhang

2022

American Journal of Physiology-Heart and Circulatory Physiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Here, we have investigated the effects of these linkers on A1 thermodynamics and function in binding to its ligand, GPIbα. Previous studies have examined the effect of the N-terminal linker or both linkers on various types of quantitative or nonquantitative binding assays and thermodynamics and found inhibitory effects on GPIbα binding from adding the N-linker or both linkers to A1 (Arce et al, 2021;Auton et al, 2012;Deng et al, 2018;Deng et al, 2017;Interlandi et al, 2017;Ju et al, 2013;Miyata & Ruggeri, 1999;Nakayama et al, 2002;Tischer et al, 2017;Tischer et al, 2014). We found that both the N and C-linker decreased A1 affinity for GPIbα and that for both the N and C-linker, O-glycosylated linkers decreased affinity more than polypeptide linkers.…”

Section: Discussionmentioning

confidence: 66%

“…It was also proposed that the N-and C-linkers interact with each other and form an autoinhibitory module that masks A1 (Deng et al, 2017). A force-dependent signature for breakage of this module has been reported that correlates with the combined expected extension of the N and C-linkers (Arce et al, 2021). An alternative mechanism has also been proposed by which the N-linker could regulate A1 affinity, i.e., by regulating the relative stability of the A1 native and intermediate states.…”

Section: Introductionmentioning

confidence: 99%

Von Willebrand factor A1 domain affinity for GPIbα and stability are differentially regulated by its O-glycosylated N-linker and C-linker

Iacob¹,

Bonazza

Hudson

et al. 2022

Preprint

View full text Add to dashboard Cite

Hemostasis in the arterial circulation is mediated by binding of the A1 domain of the ultralong protein von Willebrand factor to GPIbα on platelets to form a platelet plug. A1 is activated by tensile force on VWF concatemers imparted by hydrodynamic drag force. The A1 core is protected from force-induced unfolding by a long-range disulfide that links cysteines near its N and C-termini. The O-glycosylated linkers between A1 and its neighboring domains, which transmit tensile force to A1, are reported to regulate A1 activation for binding to GPIb, but the mechanism is controversial and incompletely defined. Here, we study how these linkers, and their polypeptide and O-glycan moieties, regulate A1 affinity by measuring affinity, kinetics, thermodynamics, hydrogen deuterium exchange (HDX), and unfolding by temperature and urea. The N-linker lowers A1 affinity 40-fold with a stronger contribution from its O-glycan than polypeptide moiety. The N-linker also decreases HDX in specific regions of A1 and increases thermal stability and the energy gap between its native state and an intermediate state, which is observed in urea-induced unfolding. The C-linker also decreases affinity of A1 for GPIbα, but in contrast to the N-linker, has no significant effect on HDX or A1 stability. Among different models for A1 activation, our data are consistent with the model that the intermediate state has high affinity for GPIbα, which is induced by tensile force physiologically and regulated allosterically by the N-linker.

show abstract

“…As seen previously for GPIbα and VWF fragments, the binding sensorgrams (Figure 2B,C) indicate this interaction proceeds with a very fast dissociation rate. 23 The kinetic fitting of such an interaction is difficult. Nevertheless, steady-state analysis of this interaction is in good agreement with ELISA results (Figure 2D).…”

Section: Asialo-aim-a1 Has Increased Binding Activitymentioning

confidence: 99%

Desialylation of O‐glycans activates von Willebrand factor by destabilizing its autoinhibitory module

Voos

Cao

Arce

et al. 2022

Journal of Thrombosis and Haemostasis

Self Cite

View full text Add to dashboard Cite

Background: The binding of the A1 domain of von Willebrand factor (VWF) to platelet receptor glycoprotein (GP)Ibα defines the VWF activity in hemostasis. Recent studies suggest that sequences flanking A1 form cooperatively an autoinhibitory module (AIM) that reduces the accessibility of the GPIbα binding site on A1. Application of a tensile force induces unfolding of the AIM. Desialylation induces spontaneous binding of plasma VWF to platelets. Most O-glycans in VWF are located around the A1 domain. Removing certain O-glycans in the flanking sequences by site-directed mutagenesis enhances A1 binding to GPIbα and produces an effect similar to type 2B von Willebrand disease in animals. Objectives: To understand if and how desialylation of O-glycans in the flanking sequences increases A1 activity. Methods: A recombinant AIM-A1 fragment encompassing VWF residues 1238-1493 and only O-glycans was treated with neuraminidase to produce desialylated protein. The glycan structure, dynamics, stability, and function of the desialylated protein was characterized by biochemical and biophysical methods and compared to the sialylated fragment.Results: Asialo-AIM-A1 exhibited increased binding activity and induced more apparent platelet aggregation than its sialylated counterpart. It exhibited a lower melting temperature, and increased hydrogen-deuterium exchange rates at residues near the secondary GPIbα binding site and the N-terminal flanking sequence. Asialo-AIM-A1 is less mechanically stable than sialo-AIM-A1, with its unstressed unfolding rate approximately 3-fold greater than the latter.Conclusions: Desialylation of O-glycans around A1 increases its activity by destabilizing the AIM.

show abstract

Activation of von Willebrand factor via mechanical unfolding of its discontinuous autoinhibitory module

Cited by 38 publications

References 88 publications

Contribution of the von Willebrand factor/ADAMTS13 imbalance to COVID-19 coagulopathy

Contribution of the von Willebrand factor/ADAMTS13 imbalance to COVID-19 coagulopathy

Von Willebrand factor A1 domain affinity for GPIbα and stability are differentially regulated by its O-glycosylated N-linker and C-linker

Desialylation of O‐glycans activates von Willebrand factor by destabilizing its autoinhibitory module

Contact Info

Product

Resources

About