Exonuclease VIII of Escherichia coli. I. Purification and physical properties

Exonuclease VIII is an enzyme whose synthesis is induced as a result of sbcA mutations. The enzyme has been purified to near homogeneity from an Escherichia coli strain containing an sbcA mutation and mutations in the structural genes for exonuclease III, exonuclease V, and endonuclease I. The enzyme specifically degrades linear duplex DNA in a reaction which requires magnesium ions and is susceptible to inhibition by other divalent cations and by sulfhydryl-blocking reagents. Enzyme activity occurs over a broad pH range with peak activity at pH 8.5 in Tris buffer. The protein has a subunit Mr = 140,000, a sedimentation coefficient of 8.4 +/- 0.6, and a Stokes radius of 142 +/- 6 A, which is consistent with its active form being a multimer. Exonuclease VIII has a frictional coefficient of 2.6 which indicates that it has an asymmetric structure.