Electrophoretic Characterization of Anti-platelet Aggregating Proteins/Peptides from Salivary Gland of Rhipicephalus (Boophilus) microplus

Ticks are economically devastating arthropod ectoparasites of livestock that exclusively feed on host blood. Ticks saliva is a crucial weapon against host-defense systems. Several small fragmented attempts are made by researchers to characterize these biomolecules. Being extremely small in size and quantity, numerous such molecules escape the deep insight of biochemists employing routine gel electrophoresis followed by Coomasie brilliant blue (CBB) staining. In a systematic manner, the present study aimed to isolate and electrophoretically characterize the salivary gland anti-platelet aggregating proteins/peptides from Rhipicephalus (Boophilus) microplus ticks. Salivary glands dissected out from female ticks were homogenized and fractionated into 120 fractions using gel filtration chromatography. All fractions were screened for anti-thrombotic activity. Nine fractions which inhibit thrombin induced platelet aggregation were electrophoretically characterized with SDS-PAGE followed by silver staining of gel. Amongst all 9 fractions which showed antiplatelet aggregating activity, protein band of approximate 64.1kDa was found predominant indicating towards peptide/protein with thrombin induced platelet aggregation inhibitory activity. Experiment was repeated many folds to evaluate the presence of the 64.1kDa bands in active fractions. Subsequently, the potential biomolecule identified need further purification and characterization to explore its therapeutic and immunization potential.