Flow molecular dynamics simulations reveal mechanosensitive regulation of von Willebrand factor through glycan-modulated autoinhibitory modules

Abstract

Force-induced protein conformational changes govern many essential biological processes, yet their molecular mechanisms remain difficult to resolve. Von Willebrand factor (VWF), a central regulator of haemostasis, is activated by hydrodynamic forces in blood flow, but how mechanical signals propagate across its multidomain architecture is poorly understood. Here, we use flow molecular dynamics (FMD), a simulation framework that applies fluid forces via controlled solvent flow to interrogate mechanosensitive proteins. Using VWF as a model system, we reconstructed the complete mechanomodule (D'D3-A1-A2-A3; 1,109 residues) with native glycosylation by integrating crystallographic data and AlphaFold predictions. FMD simulations capture a force-driven transition from a compact, autoinhibited "birds-nest" ensemble to an extended, activated state, revealing asymmetric autoinhibitory strengths within the N'AIM and C'AIM modules of the A1 domain. By directly linking static structures to dynamic, force-regulated behaviour, this work establishes a generalizable platform for dissecting protein mechanosensitivity and enabling the rational design of force-responsive therapeutics. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=48 SRC="FIGDIR/small/716521v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@edba50org.highwire.dtl.DTLVardef@1630df4org.highwire.dtl.DTLVardef@292887org.highwire.dtl.DTLVardef@23cdb7_HPS_FORMAT_FIGEXP M_FIG C_FIG Flow molecular dynamics simulations reveal that GPIb engages the A1 domain only after the disruption of key interdomain and intermodular interactions.