A Conformational Transition of the D’D3 Domain primes von Willebrand Factor for Multimerization

Abstract:

Von Willebrand factor (VWF) is a multimeric plasma glycoprotein that is critically involved in hemostasis. Biosynthesis of long VWF concatemers in the endoplasmic reticulum and the (trans-)Golgi is still not fully understood. We use the single-molecule force spectroscopy technique magnetic tweezers to analyze a previously hypothesized conformational change in the D’D3 domain crucial for VWF multimerization. We find that the interface formed by submodules C8-3, TIL3, and E3 wrapping around VWD3 can open and expose two buried cysteines, Cys1099 and Cys1142, that are vital for multimerization. By characterizing the conformational change at varying levels of force, we are able to quantify the kinetics of the transition and the stability of the interface. We find a pronounced destabilization of the interface upon lowering the pH from 7.4 to 6.2 and 5.5. This is consistent with initiation of the conformational change that enables VWF multimerization at the D’D3 domain by a decrease in pH in the trans-Golgi network and Weibel-Palade bodies. Furthermore, we find a stabilization of the interface in the presence of coagulation factor VIII (FVIII), providing evidence for a previously hypothesized binding site in submodule C8-3. Our findings highlight the critical role of the D’D3 domain in VWF biosynthesis and function and we anticipate our methodology to be applicable to study other, similar conformational changes in VWF and beyond.

SEEK ID: https://publications.h-its.org/publications/1507

DOI: 10.1182/bloodadvances.2022006978

Research Groups: Molecular Biomechanics

Publication type: Journal

Journal: Blood Advances

Citation: Blood Adv,6(17):5198–5209.

Date Published: 2nd Jun 2022

URL: https://doi.org/10.1182/bloodadvances.2022006978

Registered Mode: manually

Authors: Sophia Gruber, Achim Löf, Adina Hausch, Fabian Kutzki, Res Jöhr, Tobias Obser, Gesa König, Reinhard Schneppenheim, Camilo Aponte-Santamaría, Frauke Gräter, Maria A. Brehm, Martin Benoit, Jan Lipfert

help Submitter
Citation
Gruber, S., Löf, A., Hausch, A., Kutzki, F., Jöhr, R., Obser, T., König, G., Schneppenheim, R., Aponte-Santamaría, C., Gräter, F., Brehm, M. A., Benoit, M., & Lipfert, J. (2022). A conformational transition of the D′D3 domain primes von Willebrand factor for multimerization. In Blood Advances (Vol. 6, Issue 17, pp. 5198–5209). American Society of Hematology. https://doi.org/10.1182/bloodadvances.2022006978
Activity

Views: 2133

Created: 8th Aug 2022 at 14:37

Last updated: 5th Mar 2024 at 21:24

help Tags

This item has not yet been tagged.

help Attributions

None

Powered by
(v.1.14.2)
Copyright © 2008 - 2023 The University of Manchester and HITS gGmbH