Deoxyribonuclease II purified from the isolated lysosomes of porcine spleen and from porcine liver homogenates. Comparison with deoxyribonuclease II purified from porcine spleen homogenates.
Porcine spleen DNase II (EC 3.1.22.1), one of the best-characterized DNases II, is subcellularly located in lysosomes because the enzyme is co-sedimented with two of the lysosomal marker enzymes, cathepsin D and acid phosphatase. The physicochemical properties, including the subunit structure, sensitivity to iodoacetate inactivation, native molecular weight and chromatographic behavior, of the DNase II purified from the isolated lysosomes of porcine spleen are indistinguishable from those of the same enzyme purified from the whole porcine spleen homogenate. DNase II can also be extracted from porcine liver with 0.05 M H2SO4 or 0.1 M NaCl and purified from either extract by a series of column chromatographies. The purified liver DNase II from either extract has the same subunit structure (alpha-chain, Mr 35,000 and beta-chain, Mr 10,000) as the purified DNase II of porcine spleen. The two liver extracts as well as the extracts of spleen and gastric mucosa contain DNase II with very similar properties on Sephadex G-100 gel filtration, on acid polyacrylamide gel electrophoresis under non-denaturing conditions, and on isoelectric focusing. The data strongly suggest that, for the same species of animal, the DNase II activities in various tissues are associated with protein molecules of identical structure.
Liao
T H
TH
Department of Biochemistry, College of Medicine, National Taiwan University, Taipei, China.
Liao
W C
WC
Chang
H C
HC
Lu
K S
KS
D000818:Animals; D002458:Cell Fractionation; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D005753:Gastric Mucosa / Q000201:enzymology; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D008970:Molecular Weight; D011487:Protein Conformation; D013154:Spleen / Q000201:enzymology; D013552:Swine / Q000378:metabolism*
D007461:Iodoacetates; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II; D019807:Iodoacetic Acid