Purification and Characterization of a Chemically Induced Epstein-Barr Virus-Associated Deoxyribonuclease

Purification of Epstein-Barr virus-associated deoxyribonuclease (EBV-DNase) from Raji and P3HR-1 cells treated with 12-0-tetradecanoylphorbol-13-acetate and sodium butyrate was performed by sequential ion-exchange column chromatography and fast protein liquid chromatography. The enzyme activity, protein concentration, yield, specific activity, purification profiles, and polypeptide patterns by electrophoretic analysis in each column purification step were determined. The characteristics of the partially isolated EBV-DNase were demonstrated by the enzyme activity, DNA binding affinity, and inhibition by the nasopharyngeal carcinoma patient sera and rabbit polyclonal antibodies against the partially purified EBV-DNase. A nonisotopic assay was developed as a new method in detecting the nuclease. EBV-DNase was purified to homogeneity by FPLC. The molecular weight of the EBV-DNase was 58 KDa as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, immunostaining, and radioimmunoprecipitation using nasopharyngeal carcinoma patient sera and rabbit polyclonal antibodies