Egyptian horned viper Cerastes cerastes venom hyaluronidase: Purification, partial characterization and evidence for its action as a spreading factor

Abstract

Novel Hyaluronidase CcHaseII (33 kDa) of the most dangerous horned viper Cerastes cerastes (Cc) was purified and partial characterized in a set of biochemical assays. CcHaseII was purified by applying a protocol of two successive chromatographic steps; gel filtration on a Sephacryl S-200 and cation exchange chromatography on CM-Sepharose columns. It has specific activity 4000 units/mg protein against 154 units/mg protein for the whole venom with 26-purification fold. The enzymatic activity of the purified Hyaluronidase stimulated by Na+ and inhibited by entire tested cations, metalloproteinase inhibitors and heparin. CcHaseII (5–10 μg) enhanced one hundred percent of hemorrhagic activity of the potent purified hemorrhagic SVMP of corresponding venom (CcHTI) and enhanced edema-inducing activity of Cc venom in a dose-dependent manner. Furthermore, the described purification procedure allows simple preparation of appreciable quantities of the CcHaseII for further studies. Eventually, exploration of snake venom antigenic parts is the most crucial factor for establishing good immunogens and specific diagnostic reagents.