Purification and characterization of a neutral protease from rat-liver cytosolic fraction.
Rat liver cytosol contains a neutral protease which degrades acetylated hemoglobin and some urea-denatured proteins maximally at pH 7.5. The enzyme was purified to homogeneity by conventional chromatographic techniques. It appears to be a metalloprotease since it is inhibited by EDTA and o-phenanthroline, the metal-depleted enzyme can be reactivated by Co2+, Zn2+, Mn2+, or Mg2+, and it is not inhibited by reagents specific for carboxyl, seryl, or thiol proteases. The enzyme has an apparent molecular weight of 200,000 as determined on Sephacryl S-200 column chromatography, and electrophoresis in sodium dodecyl sulfate showed 3 protein bands corresponding to the molecular weights of 110,000, 74,000, and 40,000.
D000818:Animals; D002413:Cations, Divalent; D002845:Chromatography; D002850:Chromatography, Gel; D003600:Cytosol / Q000201:enzymology; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000302:isolation & purification*; D004789:Enzyme Activation / Q000187:drug effects; D004793:Enzyme Reactivators; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology*; D008297:Male; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D015260:Neprilysin; D010618:Phenanthrolines / Q000494:pharmacology; D011480:Protease Inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity
D002413:Cations, Divalent; D004793:Enzyme Reactivators; D008670:Metals; D010618:Phenanthrolines; D011480:Protease Inhibitors; D004492:Edetic Acid; D010450:Endopeptidases; D015260:Neprilysin; C025205:1,10-phenanthroline