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Mechano-covalent protection of coagulation factor VIII by von Willebrand factor.

Diego ButeraHaoqing Jerry WangHeng-Giap WoonYunduo Charles ZhaoLining Arnold Arnold JuPhilip Hogg
Published in: Blood advances (2022)
von Willebrand factor (VWF) is the protective carrier of pro-coagulant factor VIII (FVIII) in the shear forces of the circulation, prolonging its half-life and delivering it to the developing thrombus. Using force spectroscopy, VWF·FVIII complex formation is characterized by catch-bond behavior in which force first decelerates then accelerates bond dissociation. Patients with mutations in VWF at the FVIII binding site phenocopies hemophilia A and the most common mutations are of cysteine residues involving multiple disulfide bonds. From differential cysteine alkylation and mass spectrometry experiments, 13 VWF disulfide bonds at the FVIII binding site were found to exist in formed and unformed states, and binding of FVIII results in partial formation of 12 of the VWF bonds. Force spectroscopy studies indicate that the VWF·FVIII bond stiffens in response to force and this feature of the interaction is ablated when VWF disulfide bonds are prevented from forming, resulting in slip-only bond behavior. Exposure of VWF to pathological fluid shear forces ex vivo and in vivo causes partial cleavage of all 13 disulfide bonds, further supporting their malleable nature. These findings demonstrate that FVIII binding to VWF involves dynamic changes in the covalent states of several VWF disulfides that are required for productive interaction in physiological shear forces.
Keyphrases
  • single molecule
  • mass spectrometry
  • high resolution
  • transition metal
  • machine learning
  • deep learning
  • dna binding
  • ms ms