The Cysteine-rich Domain of Snake Venom Metalloproteinases Is a Ligand for von Willebrand Factor A Domains

The cysteine-rich domain of the haemorrhagic metalloproteinase atrolysin A was shown to inhibit collagen-stimulated platelet aggregation and to interact with MG-63 osteosarcoma cells via integrin α2β1 to inhibit adhesion to collagen I. In addition, we demonstrate by solid-phase binding assays that atrolysin A binds to collagen I and to vWF (von Willebrand factor) via exosites in the cysteine-rich domain. Interestingly, the binding site of the cysteine-rich domain on collagen I is distinct from the cell adhesion site, since the incubation of collagen-I-coated plates with the cysteine-rich domain did not prevent the adhesion of MG-63 cells to collagen. Finally, we show by surface plasmon resonance (BIAcore™) analyses that the cysteine-rich domain can block vWF binding to collagen I as well as the binding of collagen I to vWF. Taken together, these results indicate that this domain may function as a cell-surface-receptor-binding site and/or a substrate recognition exosite and may thus play a role in the pathologies associated with atrolysin A.

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Von Willebrand factor shares a distinctive cysteine-rich domain with thrombospondin and procollagen

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(87)80046-3 ◽

1987 ◽

Vol 144 (2) ◽

pp. 876-882 ◽

Cited By ~ 39

Author(s):

Lois T. Hunt ◽

Winona C. Barker

Keyword(s):

Von Willebrand Factor ◽

Rich Domain ◽

Von Willebrand ◽

Factor Shares ◽

Willebrand Factor ◽

Cysteine Rich Domain

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ADAMTS-13 cysteine-rich/spacer domains are functionally essential for von Willebrand factor cleavage

Blood ◽

10.1182/blood-2003-03-0908 ◽

2003 ◽

Vol 102 (9) ◽

pp. 3232-3237 ◽

Cited By ~ 133

Author(s):

Kenji Soejima ◽

Masanori Matsumoto ◽

Koichi Kokame ◽

Hideo Yagi ◽

Hiromichi Ishizashi ◽

...

Keyword(s):

Von Willebrand Factor ◽

Single Point ◽

Wild Type ◽

Functional Relationships ◽

Rich Domain ◽

Von Willebrand ◽

Willebrand Factor ◽

Remarkable Reduction ◽

Cysteine Rich Domain

AbstractA severe lack of von Willebrand factor–cleaving protease (VWF-CP) activity can cause thrombotic thrombocytopenic purpura (TTP). This protease was recently identified as a member of the ADAMTS family, ADAMTS-13. It consists of a preproregion, a metalloprotease domain, a disintegrin-like domain, a thrombospondin type-1 motif (Tsp1), a cysteine-rich domain, a spacer domain, additional Tsp1 repeats, and CUB domains. To explore the structural and functional relationships of ADAMTS-13, we prepared here 13 sequential COOH-terminal truncated mutants and a single-point mutant (ArgGlyAsp [RGD] to ArgGlyGlu [RGE] in the cysteine-rich domain) and compared the activity of each mutant with that of the wild-type protein. The results revealed that the truncation of the cysteine-rich/spacer domains caused a remarkable reduction in VWF-CP activity. We also prepared immunoglobulin G (IgG) fractions containing inhibitory autoantibodies against ADAMTS-13 from plasma from 3 patients with acquired TTP, and we performed mapping of their epitopes using the aforementioned mutants. The major epitopes of these antibodies were found to reside within the cysteine-rich/spacer domains. These results suggest that the ADAMTS-13 cysteine-rich/spacer domains are essential for VWF-CP activity.

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