Purification and characterization of a soluble peroxidase of rat preputial gland: comparison with lactoperoxidase.
A highly active soluble peroxidase (donor: H2O2 oxidoreductase EC 1.11.1.7) has been purified from the preputial gland of the rat by hydroxylapatite chromatography, ammonium sulfate fractionation, Sephadex gel filtration and affinity chromatography on con A-Sepharose. The enzyme shows apparent homogeneity when analysed by acid and alkaline-PAGE. Its molecular, spectral, kinetic and catalytic properties were compared with those of bovine lactoperoxidase (LPO). Preputial gland peroxidase (PPO) is a glycoprotein of molecular weight of 70-73 kDa slightly lower (78 kDa) than that of LPO. Using isoelectric focussing, PPO was resolved into eight different closely spaced protein species spanning a pI range of 5.4 to 6.4, while LPO focuses into several closely spaced protein bands in the pI range 8.5-9.3. PPO is similar to LPO in its spectral (Soret) and some kinetic properties, but it differs significantly from LPO in substrate (H2O2) tolerance and substrate inactivation. PPO also differs from LPO in showing differential inactivation by SDS. Both enzymes are glycoproteins and although concanavalin A (con A) showed a variable interaction with both enzymes, wheat germ agglutinin interacted specifically with LPO only. We suggest that PPO, the secretory peroxidase of the preputial gland, differs significantly from LPO in its molecular and catalytic properties.
De
P K
PK
Department of Physiology, Indian Institute of Chemical Biology, Calcutta.
Banerjee
R K
RK
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006139:Guaiacol / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007526:Isoelectric Point; D007700:Kinetics; D007784:Lactoperoxidase / Q000737:chemistry; D037102:Lectins; D008970:Molecular Weight; D010544:Peroxidases / Q000302:isolation & purification* / Q000378:metabolism; D011489:Protein Denaturation; D051381:Rats; D012627:Sebaceous Glands / Q000201:enzymology*; D013057:Spectrum Analysis; D014566:Urogenital System / Q000201:enzymology*
D037102:Lectins; D006139:Guaiacol; D006861:Hydrogen Peroxide; D007784:Lactoperoxidase; D010544:Peroxidases