Human urine deoxyribonuclease II (DNase II) isoenzymes: a novel immunoaffinity purification, biochemical multiplicity, genetic heterogeneity and broad distribution among tissues and body fluids.
Deoxyribonuclease II (DNase II) was purified from the urine of a 48-year-old male (a single individual) using a column chromatography series, including concanavalin A-agarose and an immunoaffinity column utilizing anti-human spleen DNase II antibody, and was then characterized. Based on the catalytic properties of the purified enzyme, we have devised a technique of isoelectric focusing by thin-layer polyacrylamide gel electrophoresis (IEF-PAGE) combined with a specific zymogram method, for investigating the possible molecular heterogeneity of human DNase II. DNase II in urine as well as the purified form was found to exist in multiple forms with different pI values separable by IEF-PAGE within a pH range of 5-7. Since sialidase treatment of the urine sample induced simplification of the isoenzyme patterns with diminishment of anodal bands, it was clear that the multiplicity of the enzyme was in part due to differences in the sialic acid content. On screening of DNase II isoenzyme patterns in urine samples from more than 200 Japanese individuals, only the common isoenzyme pattern was observed and no electrophoretic variations were detected. However, genetic studies of urinary enzyme activity and comparative studies on the activity in urine, semen and leukocytes from the same individuals suggest that the enzyme activity level of DNase II may be under genetic control. The enzyme was widely distributed in human tissues and showed high activities in secretory body fluids such as breast milk, saliva, semen and urine, and leukocyte lysates.
Yasuda
T
T
Department of Legal Medicine, Fukui Medical School, Japan.
Nadano
D
D
Awazu
S
S
Kishi
K
K
D001826:Body Fluids / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004706:Endodeoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D004996:Ethidium; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D007962:Leukocytes / Q000201:enzymology; D008297:Male; D008875:Middle Aged; D008895:Milk, Human / Q000201:enzymology; D012463:Saliva / Q000201:enzymology; D012661:Semen / Q000201:enzymology; D012685:Sepharose
D007527:Isoenzymes; D012685:Sepharose; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II; D004996:Ethidium