Gain-of-function variant p.Pro2555Arg of von Willebrand factor increases aggregate size through altering stem dynamics
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Gain-of-function variant p.Pro2555Arg of von Willebrand factor increases aggregate size through altering stem dynamics. / Huck, Volker; Chen, Po-Chia; Xu, Emma-Ruoqi; Tischer, Alexander; Klemm, Ulrike; Aponte-Santamaría, Camilo; Mess, Christian; Obser, Tobias; Kutzki, Fabian; König, Gesa; Denis, Cecile; Gräter, Frauke; Wilmanns, Matthias; Auton, Matthew; Schneider, Stefan Werner; Schneppenheim, Reinhard; Hennig, Janosch; Brehm, Maria Alexandra.
In: THROMB HAEMOSTASIS, Vol. 122, No. 2, 02.2022, p. 226-239.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Gain-of-function variant p.Pro2555Arg of von Willebrand factor increases aggregate size through altering stem dynamics
AU - Huck, Volker
AU - Chen, Po-Chia
AU - Xu, Emma-Ruoqi
AU - Tischer, Alexander
AU - Klemm, Ulrike
AU - Aponte-Santamaría, Camilo
AU - Mess, Christian
AU - Obser, Tobias
AU - Kutzki, Fabian
AU - König, Gesa
AU - Denis, Cecile
AU - Gräter, Frauke
AU - Wilmanns, Matthias
AU - Auton, Matthew
AU - Schneider, Stefan Werner
AU - Schneppenheim, Reinhard
AU - Hennig, Janosch
AU - Brehm, Maria Alexandra
N1 - The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).
PY - 2022/2
Y1 - 2022/2
N2 - The multimeric plasma glycoprotein (GP) von Willebrand factor (VWF) is best known for recruiting platelets to sites of injury during primary hemostasis. Generally, mutations in the VWF gene lead to loss of hemostatic activity and thus the bleeding disorder von Willebrand disease. By employing cone and platelet aggregometry and microfluidic assays, we uncovered a platelet GPIIb/IIIa-dependent prothrombotic gain of function (GOF) for variant p.Pro2555Arg, located in the C4 domain, leading to an increase in platelet aggregate size. We performed complementary biophysical and structural investigations using circular dichroism spectra, small-angle X-ray scattering, nuclear magnetic resonance spectroscopy, molecular dynamics simulations on the single C4 domain, and dimeric wild-type and p.Pro2555Arg constructs. C4-p.Pro2555Arg retained the overall structural conformation with minor populations of alternative conformations exhibiting increased hinge flexibility and slow conformational exchange. The dimeric protein becomes disordered and more flexible. Our data suggest that the GOF does not affect the binding affinity of the C4 domain for GPIIb/IIIa. Instead, the increased VWF dimer flexibility enhances temporal accessibility of platelet-binding sites. Using an interdisciplinary approach, we revealed that p.Pro2555Arg is the first VWF variant, which increases platelet aggregate size and shows a shear-dependent function of the VWF stem region, which can become hyperactive through mutations. Prothrombotic GOF variants of VWF are a novel concept of a VWF-associated pathomechanism of thromboembolic events, which is of general interest to vascular health but not yet considered in diagnostics. Thus, awareness should be raised for the risk they pose. Furthermore, our data implicate the C4 domain as a novel antithrombotic drug target.
AB - The multimeric plasma glycoprotein (GP) von Willebrand factor (VWF) is best known for recruiting platelets to sites of injury during primary hemostasis. Generally, mutations in the VWF gene lead to loss of hemostatic activity and thus the bleeding disorder von Willebrand disease. By employing cone and platelet aggregometry and microfluidic assays, we uncovered a platelet GPIIb/IIIa-dependent prothrombotic gain of function (GOF) for variant p.Pro2555Arg, located in the C4 domain, leading to an increase in platelet aggregate size. We performed complementary biophysical and structural investigations using circular dichroism spectra, small-angle X-ray scattering, nuclear magnetic resonance spectroscopy, molecular dynamics simulations on the single C4 domain, and dimeric wild-type and p.Pro2555Arg constructs. C4-p.Pro2555Arg retained the overall structural conformation with minor populations of alternative conformations exhibiting increased hinge flexibility and slow conformational exchange. The dimeric protein becomes disordered and more flexible. Our data suggest that the GOF does not affect the binding affinity of the C4 domain for GPIIb/IIIa. Instead, the increased VWF dimer flexibility enhances temporal accessibility of platelet-binding sites. Using an interdisciplinary approach, we revealed that p.Pro2555Arg is the first VWF variant, which increases platelet aggregate size and shows a shear-dependent function of the VWF stem region, which can become hyperactive through mutations. Prothrombotic GOF variants of VWF are a novel concept of a VWF-associated pathomechanism of thromboembolic events, which is of general interest to vascular health but not yet considered in diagnostics. Thus, awareness should be raised for the risk they pose. Furthermore, our data implicate the C4 domain as a novel antithrombotic drug target.
U2 - 10.1055/a-1344-4405
DO - 10.1055/a-1344-4405
M3 - SCORING: Journal article
C2 - 33385180
VL - 122
SP - 226
EP - 239
JO - THROMB HAEMOSTASIS
JF - THROMB HAEMOSTASIS
SN - 0340-6245
IS - 2
ER -