title: [Catalytic and immunochemical properties of ferritin conjugates with horseradish peroxidase].
abstract: The human spleen ferritin--horseradish peroxidase conjugate (HRP--Fer) was synthesized by periodate oxidation of the enzyme carbohydrate fragment. The protein fraction containing 1-2 peroxidase molecules and characterized by kinetic homogeneity was obtained in the peroxidatic ortho-dianisidine (o-DA) oxidation reaction. Gel diffusion precipitation of HRP--Fer with peroxidases and ferritin antibodies was carried out. The precipitation confirms the retention by peroxidase and ferritin of their antigenic properties. The kinetics of peroxidatic oxidation of o-DA by the HRP--Fer conjugate was studied within the temperature interval of 15-37 degrees C. The value of catalytic constant for this reaction exceeds that for native peroxidase 1.75-fold. A kinetic analysis of thermal inactivation of peroxidase and its conjugate was performed within the temperature range of 40-65 degrees C. The effective rate constants of inactivation obtained from the first order equation are higher for HRP--Fer than for the native enzyme. The effect of pH on the rates of inactivation of HRP--Fer and the non-modified enzyme was studied at 50 degrees C. The enzyme and its conjugate were shown to stabilize in acid media. The HRP--Fer conjugate can be used as an effective tool in immunoenzymatic assays of ferritin.
authors.forename: V N
authors.initials: VN
authors.lastname: Denisov
authors.forename: D I
authors.initials: DI
authors.lastname: Metelitsa
mesh_terms: D002384:Catalysis; D005293:Ferritins / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D007700:Kinetics; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism
chemical_list: D005293:Ferritins; D006735:Horseradish Peroxidase; D010544:Peroxidases
vernacular_title: Kataliticheskie i immunokhimicheskie svoĭstva kon''iugata ferritina s peroksidazoĭ iz khrena.