Hepatic Nucleases. 1. Methods for the Specific Determination and Characterization in Rat Liver

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Abstract
With a view to the study of the subcellular localization of nucleases, methods ensuring the specificity of enzyme determinations are presented for six types of nuclease activities in liver homogenates. The ribonuclease activity of rat liver is due to three enzymes with different pH optimum. For acid ribonuclease (pH optimum 5.3), it is possible to avoid interference from the other ribonucleases by performing the incubation at pH 5. Neutral ribonuclease (pH optimum 7.6) is differentiated by relying on its sensitivity to the natural inhibitor from the supernatant of liver homogenate. Comparison of activities before and after pretreatment at 50 °C in acid medium permits the specific measurement of alkaline ribonuclease (pH optimum 8.8). The optimal conditions for the determination in liver homogenates of two deoxyribonucleases and of an enzyme acting on polyriboadenylate are also described. The activity of these various nucleases is compared and some of their properties are investigated.

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