Immobilized betulinic acid column and its interactions with phospholipase A2 and snake venom proteins

Betulinic acid (BA) is a plant-derived pentacyclic triterpenoid. Although BA has been found to have diverse pharmacological effects, including anti-tumor and anti-inflammatory actions and potential as inhibitor of phospholipase A2 (PLA2), its cellular targets remain unclear. In this study, BA was immobilized onto an acrylamide matrix. The immobilized-BA column could retain the purified PLA2 of bovine pancreas or the PLA2 of snake venom from Naja nigricollis. The bound PLA2 were not eluted by high salt concentrations but were eluted by either acid or calcium free buffer. Besides the PLA2, a group of basic proteins of snake venom with molecular weights of about 7 kDa were also strongly bound by immobilized BA. One of these proteins was identified as gamma-cardiotoxin. The usefulness of immobilized BA for exploring the cellular targets of BA is discussed.