Domain-specific mechanical modulation of VWF-ADAMTS13 interaction

Mol Biol Cell. 2019 Jul 22;30(16):1920-1929. doi: 10.1091/mbc.E19-01-0021. Epub 2019 May 8.


Hemodynamic forces activate the Von Willebrand factor (VWF) and facilitate its cleavage by a disintegrin and metalloprotease with thrombospondin motifs-13 (ADAMTS13), reducing the adhesive activity of VWF. Biochemical assays have mapped the binding sites on both molecules. However, these assays require incubation of two molecules for a period beyond the time allowed in flowing blood. We used a single-molecule technique to examine these rapid, transient, and mechanically modulated molecular interactions in short times under forces to mimic what happens in circulation. Wild-type ADAMTS13 and two truncation variants that either lacked the C-terminal thrombospondin motif-7 to the CUB domain (MP-TSP6) or contained only the two CUB domains (CUB) were characterized for interactions with coiled VWF, flow-elongated VWF, and a VWF A1A2A3 tridomain. These interactions exhibited distinctive patterns of calcium dependency, binding affinity, and force-regulated lifetime. The results suggest that 1) ADAMTS13 binds coiled VWF primarily through CUB in a calcium-dependent manner via a site(s) outside A1A2A3, 2) ADAMTS13 binds flow-extended VWF predominantly through MP-TSP6 via a site(s) different from the one(s) at A1A2A3; and 3) ADAMTS13 binds A1A2A3 through MP-TSP6 in a Ca2+-dependent manner to autoinhibit another Ca2+-independent binding site on CUB. These data reveal that multiple sites on both molecules are involved in mechanically modulated VWF-ADAMTS13 interaction.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAMTS13 Protein / chemistry*
  • ADAMTS13 Protein / metabolism*
  • Binding Sites
  • Biomechanical Phenomena
  • Cations
  • HEK293 Cells
  • Humans
  • Kinetics
  • Protein Binding
  • Protein Domains
  • von Willebrand Factor / chemistry*
  • von Willebrand Factor / metabolism*


  • Cations
  • von Willebrand Factor
  • ADAMTS13 Protein