Integrins are a family of heterodimeric transmembrane receptors which link the extracellular matrix to the cell cytoskeleton. These receptors play a role in many cellular processes: adhesion, proliferation, migration, apoptosis, and platelet aggregation, thus modulating a wide range of scenarios in health and disease. Therefore,
integrins have been the target of new antithrombotic drugs.
Disintegrins from
snake venoms are recognized by the ability to modulate the activity of
integrins, such as
integrin αIIbβ3, a fundamental
platelet glycoprotein, and αvβ3 expressed on
tumor cells. For this reason,
disintegrins are unique and potential tools for examining
integrin-matrix interaction and the development of novel
antithrombotic agents. The present study aims to obtain the recombinant form of jararacin and evaluate the secondary structure and its effects on hemostasis and
thrombosis. rJararacin was expressed in the Pichia pastoris (P. pastoris) expression system and purified the
recombinant protein with a yield of 40 mg/L of culture. The molecular mass (7722 Da) and internal sequence were confirmed by mass spectrometry. Structure and folding analysis were obtained by Circular Dichroism and 1H Nuclear Magnetic Resonance spectra.
Disintegrin structure reveals properly folded with the presence of β-sheet structure. rJararacin significantly demonstrated inhibition of the adhesion of B16F10 cells and platelets to the
fibronectin matrix under static conditions. rJararacin inhibited platelet aggregation induced by
ADP (IC50 95 nM),
collagen (IC50 57 nM), and
thrombin (IC50 22 nM) in a dose-dependent manner. This
disintegrin also inhibited 81% and 94% of the adhesion of platelets to
fibrinogen and
collagen under continuous flow, respectively. In addition, rjararacin efficaciously prevents platelet aggregation in vitro and ex vivo with rat platelets and
thrombus occlusion at an effective dose (5 mg/kg). The data here provides evidence that rjararacin possesses the potential as an αIIbβ3 antagonist, capable of preventing arterial
thrombosis.