Identification of sites in the cysteine-rich domain of the class P-III snake venom metalloproteinases responsible for inhibition of platelet function

• Published in: FEBS letters Vol. 549; no. 1; pp. 129 - 134
• Main Authors: Kamiguti, Aura S., Gallagher, Paul, Marcinkiewicz, Cezary, Theakston, R.David G., Zuzel, Mirko, Fox, Jay W.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 14.08.2003
• Subjects: Amino Acid Sequence; Animals; Bothrops jararaca Venom; Cell Adhesion - drug effects; CMFDA, 5-chloromethyl fluorescein diacetate; Collagen - drug effects; Crotalid Venoms - chemistry; Crotalid Venoms - pharmacology; Cysteine; Cysteine-rich domain; GPVI, glycoprotein VI; HBSS, Hank's balanced salt solution; Humans; Integrin alpha2 - metabolism; K562 Cells; Metalloendopeptidases - chemistry; Metalloendopeptidases - pharmacology; PGE1, prostaglandin E1; Platelet Activation - drug effects; Platelet aggregation; Platelet Aggregation Inhibitors - chemistry; Platelet Aggregation Inhibitors - pharmacology; Protein Structure, Tertiary; Protein tyrosine phophorylation; PRP, platelet-rich plasma; RIPA, radioimmunoprecipitation assay; Snake venom metalloproteinase; Snake Venoms - enzymology; SVMP, snake venom metalloproteinase; α2β1 integrin; SVMP, snake venom metalloproteinase; PGE1, prostaglandin E1; PRP, platelet-rich plasma; Cysteine-rich domain; CMFDA, 5-chloromethyl fluorescein diacetate; α2β1 integrin; HBSS, Hank’s balanced salt solution; RIPA, radioimmunoprecipitation assay; Snake venom metalloproteinase; Platelet aggregation; GPVI, glycoprotein VI; Protein tyrosine phophorylation
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(03)00799-3

Atrolysin A and jararhagin are class P-III snake venom metalloproteinases (SVMPs) with three distinct domains: a metalloproteinase, a disintegrin-like and a cysteine-rich. The metalloproteinase and the disintegrin-like domains of atrolysin A and jararhagin contain peptide sequences that interact with α2β1 integrin and inhibit the platelet responses to collagen. Recently, the recombinant cysteine-rich domain of atrolysin A was shown to have similar effects, but the sequence(s) responsible for this is unknown. In this report, we demonstrate two complete peptide sequences from the homologous cysteine-rich domains of atrolysin A and jararhagin that inhibit both platelet aggregation by collagen and adhesion of α2-expressing K562 cells to this protein. In addition, the peptide effects on platelets do not seem to involve an inhibition of GPVI. These results identify, for the first time, sites in the cysteine-rich domain of SVMPs that inhibit cell responses to collagen and reveal the complexity of the potential biological effects of these enzymes with multifunctional domains.