Further purification and properties of rat uterine peroxidase.
Peroxidase was purified 3700-fold from homogenates of estradiol-treated rat uteri by affinity chromatography on concanavalin A (ConA)- Sepharose followed by gel filtration on Bio-Gel P-150 with high recovery of enzyme. A single protein (molecular weight (MW) 45 000) staining for heme was shown by sodium dodecyl sulfate - polyacrylamide gel electrophoresis to be present in the peak fractions of enzymic activity eluted from the ConA-Sepharose column. This protein had the same mobility as bovine lactoperoxidase (MW 78 000) in a cationic gel electrophoretic system under nondenaturing conditions. Peroxidase activity in a NaCl extract of the uterus was lower than that in a CaCl2 extract but was unaffected by prolonged storage at - 20 degrees C. In contrast, the CaCl2-extracted enzyme lost much of its activity under these conditions by a process which could by prevented by the addition of glycerol. The sulfhydryl reagent, N-ethylmaleimide, which caused a marked increase in the activity of uterine peroxidase, provided only partial protection against inactivation during storage of CaCl2 extracts of this enzyme at low temperature.
D000818:Animals; D002122:Calcium Chloride; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D008970:Molecular Weight; D010544:Peroxidases / Q000302:isolation & purification*; D051381:Rats; D014599:Uterus / Q000201:enzymology*
D010544:Peroxidases; D002122:Calcium Chloride; D005033:Ethylmaleimide; D005990:Glycerol