title: Function of the cysteine-rich domain of the haemorrhagic metalloproteinase atrolysin A: targeting adhesion proteins collagen I and von Willebrand factor.
abstract: 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 alpha2beta1 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.
authors.affiliation: Laboratório Especial de Toxinologia Aplicada and CAT-CEPID, Instituto Butantan, Sao Paulo, SP 05503-900, Brazil. solangeserrano@butantan.gov.br
authors.forename: Solange M T
authors.initials: SM
authors.lastname: Serrano
authors.forename: Li-Guo
authors.initials: LG
authors.lastname: Jia
authors.forename: Deyu
authors.initials: D
authors.lastname: Wang
authors.forename: John D
authors.initials: JD
authors.lastname: Shannon
authors.forename: Jay W
authors.initials: JW
authors.lastname: Fox
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins; D012910:Snake Venoms / Q000737:chemistry; D020349:Surface Plasmon Resonance; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D024042:Collagen Type I; D011994:Recombinant Proteins; D012910:Snake Venoms; D014841:von Willebrand Factor; D008666:Metalloendopeptidases; C044577:atrolysin A; D003545:Cysteine