TY - JOUR
T1 - Purification and properties of a new DNase activity from KB cells
AU - Frenkel, Gerald D.
AU - Randies, Karen
AU - Berns, Neil
N1 - Funding Information: This work was supported in part by Grant CA28084 from the National Cancer Funding Information: Institute and Grant NP-198 from the American Cancer Society.
PY - 1981/12/11
Y1 - 1981/12/11
N2 - A deoxyribonuclease activity with specificity towards single-stranded DNA has been purified approximately four hundred-fold from KB cells, by chromatography on DEAE-cellulose, phosphocellulose and hydroxylapatite. The last step of the purification results in separation of the enzyme from a DNase activity which has been described previously (Wang, E.C., Furth, J.J. and Rose, J.A., (1978) Biochemistry 17: 544-549). The properties of the new DNase activity are significantly different from those of the enzymes which have previously been identified in these cells. The activity sediments at approximately 7.5S in a glycerol gradient. The DNase activity is optimal at pHs between 6.0 and 6.5. It cleaves DNA endonucleolytically and hydrolyzes single-stranded DNA at about 11 times the rate of double-stranded DNA and at twice the rate of Poly (dA). The activity is moderately sensitive to inhibition by N-ethylmaleimide and is inhibited 80% by 50 mM NaCl. It is stimulated twenty-fold by Mn++ at an optimal concentration of approximately 0.7 mM. It is stimulated to a lesser extent by Mg++, but not by Ca++.
AB - A deoxyribonuclease activity with specificity towards single-stranded DNA has been purified approximately four hundred-fold from KB cells, by chromatography on DEAE-cellulose, phosphocellulose and hydroxylapatite. The last step of the purification results in separation of the enzyme from a DNase activity which has been described previously (Wang, E.C., Furth, J.J. and Rose, J.A., (1978) Biochemistry 17: 544-549). The properties of the new DNase activity are significantly different from those of the enzymes which have previously been identified in these cells. The activity sediments at approximately 7.5S in a glycerol gradient. The DNase activity is optimal at pHs between 6.0 and 6.5. It cleaves DNA endonucleolytically and hydrolyzes single-stranded DNA at about 11 times the rate of double-stranded DNA and at twice the rate of Poly (dA). The activity is moderately sensitive to inhibition by N-ethylmaleimide and is inhibited 80% by 50 mM NaCl. It is stimulated twenty-fold by Mn++ at an optimal concentration of approximately 0.7 mM. It is stimulated to a lesser extent by Mg++, but not by Ca++.
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U2 - https://doi.org/10.1093/nar/9.23.6635
DO - https://doi.org/10.1093/nar/9.23.6635
M3 - Article
C2 - 7322924
VL - 9
SP - 6635
EP - 6644
JO - Nucleic acids research
JF - Nucleic acids research
SN - 0305-1048
IS - 23
ER -