Hemostatic effects of recombinant DisBa-01, a disintegrin from Bothrops alternatus.
A monomeric RGD-disintegrin was recently identified from a cDNA library from the venom gland of Bothrops alternatus. The corresponding 12 kDa-recombinant protein, DisBa-01, specifically interacted with alpha(v)beta3 integrin and displayed potent anti-metastatic and anti-angiogenic properties. Here, the interaction of DisBa-01 with platelet alphaIIb beta3 integrin and its effects on hemostasis and thrombosis were investigated. DisBa-01 bound to Chinese Hamster Ovary (CHO) cells expressing beta3 or alphaIIb beta3 and promoted their adhesion and the adhesion of resting platelets onto glass coverslips. The disintegrin inhibited the binding of FITC-fibrinogen and FITC-PAC-1 to ADP-stimulated platelets and inhibited ADP-, TRAP- and collagen-induced aggregation of murine, rabbit or human platelets. In a flow chamber assay, DisBa-01 inhibited and reverted platelet adhesion to immobilized fibrinogen. DisBa-01 inhibited the phosphorylation of FAK following platelet activation. The intravenous injection of DisBa-01 in C57Bl6/j mice, prolonged tail bleeding time as well as thrombotic occlusion time in mesenteric venules and arterioles following vessel injury with FeCl3. In conclusion, DisBa-01 antagonizes the platelet alphaIIb beta3 integrin and potently inhibits thrombosis.
INSERM U553, Paris, F-75475 France.
Alexandre
A
Kauskot
Marcia R
MR
Cominetti
Oscar H P
OH
Ramos
Iga
I
Bechyne
Jean-Marie
JM
Renard
Marc F
MF
Hoylaerts
Michel
M
Crepin
Chantal
C
Legrand
Heloisa S
HS
Selistre-de-Araujo
Arnaud
A
Bonnefoy
D000818:Animals; D017837:Bothrops; D016466:CHO Cells / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D003435:Crotalid Venoms / Q000633:toxicity*; D051417:Focal Adhesion Kinase 1 / Q000187:drug effects / Q000378:metabolism; D006487:Hemostasis / Q000187:drug effects*; D006490:Hemostatics / Q000494:pharmacology*; D006801:Humans; D010766:Phosphorylation; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D011994:Recombinant Proteins / Q000633:toxicity; D014162:Transfection
D003435:Crotalid Venoms; D006490:Hemostatics; D011994:Recombinant Proteins; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human