ADAMTS-13 metalloprotease interacts with the endothelial cell-derived ultra-large von Willebrand factor.
Thrombotic thrombocytopenic purpura is caused by congenital or acquired deficiency of ADAMTS-13, a metalloprotease that cleaves the endothelium-derived ultra-large multimers of von Willebrand factor (ULVWF). The proteolysis converts hyper-reactive and thrombogenic ULVWF into smaller and less adhesive plasma forms. Activity of ADAMTS-13 is usually measured in a static system under non-physiological conditions that require protein denaturation and prolonged incubation. We have demonstrated previously that ULVWF multimers, upon release from endothelial cells, form platelet-decorated string-like structures that are rapidly cleaved by ADAMTS-13. Here we report the direct interaction between ADAMTS-13 and VWF under both static and flowing conditions. ADAMTS-13-coated beads adhered to both immobilized VWF and ULVWF strings presented by stimulated endothelial cells. These beads adhered to VWF under both venous (2.5 dynes/cm2) and arterial (30 dynes/cm2) shear stresses. We then demonstrated that ADAMTS-13 beads adhered to immobilized recombinant VWF-A1 and -A3 domains, but soluble metalloprotease bound preferentially to the A3 domain, suggesting that the VWF A3 domain may be the primary docking site for the metalloprotease. We suggest that tensile stresses imposed by fluid shear stretch endothelial bound ULVWF multimers to expose binding sites within the A domains for circulating ADAMTS-13. The bound enzyme then cleaves within the A2 domain that lies in close proximity and releases smaller VWF multimers into the plasma. Once released, these cleaved VWF fragments become inaccessible for the metalloprotease to prevent further cleavage.
Dong
Jing-fei
JF
Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, USA. jfdong@bcm.tmc.edu
Moake
Joel L
JL
Bernardo
Aubrey
A
Fujikawa
Kazuo
K
Ball
Chalmette
C
Nolasco
Leticia
L
López
José A
JA
Cruz
Miguel A
MA
D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D009154:Mutation; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000302:isolation & purification; D011137:Polystyrenes / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology; D014841:von Willebrand Factor / Q000737:chemistry / Q000378:metabolism*
D019038:Platelet Glycoprotein GPIb-IX Complex; D011137:Polystyrenes; D011994:Recombinant Proteins; C023592:glycocalicin; D014841:von Willebrand Factor; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human