Haemolytic activity of complement component C3 of Labeo rohita
Pushpa Choudhary, M Makesh and Jitendra Kumar
Complement component C3 is one of the most abundant complement components found in the plasma/serum of animals and which participates in all three complement activation pathways. In the present study Complement component C3 of rohu, Labeo rohita was purified from plasma of rohu by two step column chromatography viz, anion and cation exchange chromatography. The purified rohu C3 protein showed the characteristic two chain structure, α and β in reducing SDS-PAGE. The molecular weight of α and β chain was found to be 112 kDa and 77 kDa respectively while in non reducing SDS-PAGE it showed a single band with molecular weight of 189 kDA. The assessment of haemolytic activity of purified rohu C3 was determined by alternative complement pathway and classical complement pathway. Zymosan treated rohu serum showed negligible hemolysis of sheep RBCs while complete hemolysis was observed by addition of purified rohu C3. Dose dependent hemolysis of sheep RBCs was observed in alternative complement pathway as well as in classical complement pathway.