Publications

BibTex format

@article{O'Brien:2021:10.1111/jth.15142,
author = {O'Brien, HER and Zhang, XF and Sanz-Hernandez, M and Chion, A and Shapiro, S and Mobayen, G and Xu, Y and De, Simone A and Laffan, MA and McKinnon, TAJ},
doi = {10.1111/jth.15142},
journal = {Journal of Thrombosis and Haemostasis},
pages = {358--369},
title = {Blocking von Willebrand factor free thiols inhibits binding to collagen under high and pathological shear stress},
url = {http://dx.doi.org/10.1111/jth.15142},
volume = {19},
year = {2021}
}

Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - BackgroundVon Willebrand factor (VWF) contains a number of free thiols, the majority of which are located in its Cdomains, and these have been shown to alter VWF function, However, the impact of free thiols on function following acute exposure of VWF to collagen under high and pathological shear stress has not been determined.MethodsVWF free thiols were blocked with Nethylmaleimide and flow assays performed under high and pathological shear rates to determine the impact on platelet capture and collagen binding function. Atomic force microscopy (AFM) was used to probe the interaction of VWF with collagen and molecular simulations conducted to determine the effect of free thiols on the flexibility of the VWFC4 domain.ResultsBlockade of VWF free thiols reduced VWFmediated platelet capture to collagen in a sheardependent manner, with platelet capture virtually abolished above 5000 s−1 and in regions of stenosis in microfluidic channels. Direct visualization of VWF fibers formed under extreme pathological shear rates and analysis of collagenbound VWF attributed the effect to altered binding of VWF to collagen. AFM measurements showed that thiolblockade reduced the lifetime and strength of the VWFcollagen bond. Pulling simulations of the VWFC4 domain demonstrated that with one or two reduced disulphide bonds the C4 domain has increased flexibility and the propensity to undergo freethiol exchange.ConclusionsWe conclude that free thiols in the Cdomains of VWF enhance the flexibility of the molecule and enable it to withstand high shear forces following collagen binding, demonstrating a previously unrecognized role for VWF free thiols.
AU  - O'Brien,HER
AU  - Zhang,XF
AU  - Sanz-Hernandez,M
AU  - Chion,A
AU  - Shapiro,S
AU  - Mobayen,G
AU  - Xu,Y
AU  - De,Simone A
AU  - Laffan,MA
AU  - McKinnon,TAJ
DO  - 10.1111/jth.15142
EP  - 369
PY  - 2021///
SN  - 1538-7836
SP  - 358
TI  - Blocking von Willebrand factor free thiols inhibits binding to collagen under high and pathological shear stress
T2  - Journal of Thrombosis and Haemostasis
UR  - http://dx.doi.org/10.1111/jth.15142
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000590166400001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - https://onlinelibrary.wiley.com/doi/10.1111/jth.15142
UR  - http://hdl.handle.net/10044/1/85403
VL  - 19
ER  -

Download

ProfessorMikeLaffan

Contact

Assistant

Location

Summary

Citation

BibTex format

RIS format (EndNote, RefMan)