Two deoxyribonucleases from Novikoff ascites hepatoma cells

Publisher Website
Google Scholar
Abstract
Two DNases have been isolated and separated from Novikoff ascites hepatoma by ammonium sulfate fractionation and have been further purified by chromatography on ion-exchange columns of DEAE types.The partially purified acid DNase is free of any measurable RNase activity, while the partially purified alkaline DNase preparation still exhibits RNase activity.The alkaline DNase requires sulfhydryl compounds for maximum activity, whereas the acid DNase does not. Both DNases require Mg2+ ions for maximum activity. EDTA strongly inhibits the alkaline DNase activity and the inhibition can be reversed by the addition of Mg2+ ions. On the other hand, EDTA activates the acid DNase either in the presence or in the absence of Mg2+.Sarkomycin inhibits the alkaline DNase but does not inhibit the acid DNase. Actinomycin D and heparin inhibit both DNase activities.The products of the alkaline DNase digestion consist of four deoxymononucleotides as well as higher oligonucleotides, all terminating in 5′-phosphate. The alkaline DNase seems to exhibit an endonucleolytic mode of attack in the early stage of hydrolysis with a subsequent exonucleolytic action. However, the possibility of contamination by an unknown exonuclease cannot be ruled out. On the other hand, the products of the acid DNase digestion consist mainly of oligonucleotides with average chain length larger than 8 units all terminating in 3′-phosphate. No mononucleotides can be detected. This suggests that the acid DNase is a typical endonuclease and possesses no detectable exonuclease activity.The acid DNase preferentially attacks linkages of the type dPupGp, whereas the preferential linkage(s) for the alkaline DNase has not been established.

This publication has 0 references indexed in Scilit: