title: Human urine DNase I: immunological identity with human pancreatic DNase I, and enzymic and proteochemical properties of the enzyme.
abstract: DNase I in human urine was purified to an electrophoretically homogeneous state by column chromatographies on DEAE-lignocellulose, hydroxyapatite, DEAE-cellulose, Sephadex G-75 and elastin-celite. The purified enzyme was immunologically identical with human pancreatic DNase I, but not with bovine pancreatic DNase I. The molecular weight and isoelectric point of the enzyme were estimated to be 4.1 X 10(4) and 3.6, respectively. The amino acid analysis revealed that 1 mol of the enzyme contained 8 mol of half-cystine. The N-terminal amino acid was identified as leucine by the dansyl chloride method. The enzyme was active in the presence of Mg2+, Co2+, or Mn2+, The optimum pH was around 6.5. The enzyme was stable in the pH range from 5.0 to 9.0 and at temperatures lower than 45 degrees C. The rate of hydrolysis of native DNA by the enzyme was twice as fast as that observed with heat-denatured DNA. This enzyme exhaustively degraded about 20% of the phosphodiester bonds in native DNA. The enzyme also degraded poly(dA) and poly(dT), but hardly degraded poly(dG) and poly(dC).
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D003850:Deoxyribonuclease I; D004706:Endodeoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007700:Kinetics; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology*; D013045:Species Specificity
chemical_list: D000596:Amino Acids; D002413:Cations, Divalent; D000939:Epitopes; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I