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Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers

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Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers. / Lippok, Svenja; Radtke, Matthias; Obser, Tobias; Kleemeier, Lars; Schneppenheim, Reinhard; Budde, Ulrich; Netz, Roland R; Rädler, Joachim O.

In: BIOPHYS J, Vol. 110, No. 3, 02.02.2016, p. 545-54.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Lippok, S, Radtke, M, Obser, T, Kleemeier, L, Schneppenheim, R, Budde, U, Netz, RR & Rädler, JO 2016, 'Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers', BIOPHYS J, vol. 110, no. 3, pp. 545-54. https://doi.org/10.1016/j.bpj.2015.12.023

APA

Lippok, S., Radtke, M., Obser, T., Kleemeier, L., Schneppenheim, R., Budde, U., Netz, R. R., & Rädler, J. O. (2016). Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers. BIOPHYS J, 110(3), 545-54. https://doi.org/10.1016/j.bpj.2015.12.023

Vancouver

Lippok S, Radtke M, Obser T, Kleemeier L, Schneppenheim R, Budde U et al. Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers. BIOPHYS J. 2016 Feb 2;110(3):545-54. https://doi.org/10.1016/j.bpj.2015.12.023

Bibtex

@article{6ddec18c42104bfb8e176aaca9858d66,
title = "Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers",
abstract = "Proteolysis of the multimeric blood coagulation protein von Willebrand Factor (VWF) by ADAMTS13 is crucial for prevention of microvascular thrombosis. ADAMTS13 cleaves VWF within the mechanosensitive A2 domain, which is believed to open under shear flow. In this study, we combine fluorescence correlation spectroscopy (FCS) and a microfluidic shear cell to monitor real-time kinetics of full-length VWF proteolysis as a function of shear stress. For comparison, we also measure the Michaelis-Menten kinetics of ADAMTS13 cleavage of wild-type VWF in the absence of shear but partially denaturing conditions. Under shear, ADAMTS13 activity on full-length VWF arises without denaturing agent as evidenced by FCS and gel-based multimer analysis. In agreement with Brownian hydrodynamics simulations, we find a sigmoidal increase of the enzymatic rate as a function of shear at a threshold shear rate γ˙1/2 = 5522/s. The same flow-rate dependence of ADAMTS13 activity we also observe in blood plasma, which is relevant to predict hemostatic dysfunction.",
author = "Svenja Lippok and Matthias Radtke and Tobias Obser and Lars Kleemeier and Reinhard Schneppenheim and Ulrich Budde and Netz, {Roland R} and R{\"a}dler, {Joachim O}",
note = "Copyright {\textcopyright} 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.",
year = "2016",
month = feb,
day = "2",
doi = "10.1016/j.bpj.2015.12.023",
language = "English",
volume = "110",
pages = "545--54",
journal = "BIOPHYS J",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "3",

}

RIS

TY - JOUR

T1 - Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers

AU - Lippok, Svenja

AU - Radtke, Matthias

AU - Obser, Tobias

AU - Kleemeier, Lars

AU - Schneppenheim, Reinhard

AU - Budde, Ulrich

AU - Netz, Roland R

AU - Rädler, Joachim O

N1 - Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

PY - 2016/2/2

Y1 - 2016/2/2

N2 - Proteolysis of the multimeric blood coagulation protein von Willebrand Factor (VWF) by ADAMTS13 is crucial for prevention of microvascular thrombosis. ADAMTS13 cleaves VWF within the mechanosensitive A2 domain, which is believed to open under shear flow. In this study, we combine fluorescence correlation spectroscopy (FCS) and a microfluidic shear cell to monitor real-time kinetics of full-length VWF proteolysis as a function of shear stress. For comparison, we also measure the Michaelis-Menten kinetics of ADAMTS13 cleavage of wild-type VWF in the absence of shear but partially denaturing conditions. Under shear, ADAMTS13 activity on full-length VWF arises without denaturing agent as evidenced by FCS and gel-based multimer analysis. In agreement with Brownian hydrodynamics simulations, we find a sigmoidal increase of the enzymatic rate as a function of shear at a threshold shear rate γ˙1/2 = 5522/s. The same flow-rate dependence of ADAMTS13 activity we also observe in blood plasma, which is relevant to predict hemostatic dysfunction.

AB - Proteolysis of the multimeric blood coagulation protein von Willebrand Factor (VWF) by ADAMTS13 is crucial for prevention of microvascular thrombosis. ADAMTS13 cleaves VWF within the mechanosensitive A2 domain, which is believed to open under shear flow. In this study, we combine fluorescence correlation spectroscopy (FCS) and a microfluidic shear cell to monitor real-time kinetics of full-length VWF proteolysis as a function of shear stress. For comparison, we also measure the Michaelis-Menten kinetics of ADAMTS13 cleavage of wild-type VWF in the absence of shear but partially denaturing conditions. Under shear, ADAMTS13 activity on full-length VWF arises without denaturing agent as evidenced by FCS and gel-based multimer analysis. In agreement with Brownian hydrodynamics simulations, we find a sigmoidal increase of the enzymatic rate as a function of shear at a threshold shear rate γ˙1/2 = 5522/s. The same flow-rate dependence of ADAMTS13 activity we also observe in blood plasma, which is relevant to predict hemostatic dysfunction.

U2 - 10.1016/j.bpj.2015.12.023

DO - 10.1016/j.bpj.2015.12.023

M3 - SCORING: Journal article

C2 - 26840720

VL - 110

SP - 545

EP - 554

JO - BIOPHYS J

JF - BIOPHYS J

SN - 0006-3495

IS - 3

ER -