PURIFICATION AND CHARACTERIZATION OF C-TYPE LECTIN FROM BOTHROPS LEUCURUS SNAKE VENOM WITH ANTI-PLATELET AND ANTI-METASTATIC POTENTIAL

Introduction: Snake venom lectins are categorized into classical sugar-binding C-type lectins and non-sugar binding C-type-like lectins. C-type lectins are typically homodimeric proteins with affinity for beta-galactoside residues and are known to exhibit diverse biological activities, including the inhibition of platelet aggregation. Platelets play a crucial role in several physiological and pathological processes, such as thrombosis and metastasis. This study aimed to elucidate the biochemical and functional properties of leuccetin, a C-type lectin isolated from Bothrops leucurus venom, with a particular focus on its effects on platelet function.

Methods: Leuccetin was purified via affinity chromatography using D-galactose gel resin. Its effects on platelet function were assessed using washed platelets and platelet-rich plasma. The potential anti-metastatic activity of leuccetin was evaluated using MDA-MB-231 breast cancer cells. Platelet–tumor cell interactions were quantified using a plate reader-based assay.

Results: Leuccetin displayed a molecular mass of approximately 15 kDa under reducing conditions and 30 kDa under non-reducing conditions. Functionally, leucceytin inhibits platelet aggregation induced by convulxin (CVX) and the von Willebrand factor (vWF). Western Blot analysis demonstrated that leuccetin binds to key platelet receptors—GPVI, GPIb, αV integrin, and CLEC-2—correlating with its inhibitory effects on platelet aggregation. Notably, leuccetin also inhibited platelet aggregation induced by MDA-MB-231 and reduced the adhesion of these cells at non-cytotoxic concentrations. Furthermore, leuccetin disrupted platelet–tumor cell interaction.

Conclusion: These findings suggest that leuccetin has biotechnological potential as a novel antiplatelet and/or anti-metastatic agent, due to its ability to inhibit platelet aggregation and platelet–tumor cell interactions.