Role of the apoprotein in the catalytic peroxidase-like function of ferritin.
The thermal inactivation of horse spleen ferritin was studied over a range of temperatures (36-52 degrees C) in 0.1 M acetate buffer (pH 4.2) as a decrease of its peroxidase activity during tetramethylbenzidine (TMB) oxidation by hydrogen peroxide. The activation energy of this process was 163.3 kJ/mole. Thermodynamic activation parameters for the loss of peroxidase activity of ferritin were calculated. The influence of various detergents on ferritin-dependent oxidation of TMB, ortho-tolidine, and ortho-phenylenediamine (PDA) by hydrogen peroxide was studied in 0.1 M phosphate buffer (pH 6.0) at 20 degrees C. Relatively high concentrations of charged detergents (SDS and cetyltrimethylammonium bromide) decreased the peroxidase activity of ferritin with all three amines, whereas moderate concentrations of the nonionic detergent Triton X-100 did not influence oxidation of these substrates. Increase of dimethylformamide concentration in 0.02 M acetate buffer (pH 4.2) from 5 to 40% strongly decreased the rate of TMB and PDA oxidation by hydrogen peroxide or cumene hydroperoxide. Decrease in the activity of thermally inactivated ferritin with TMB as substrate, reduction of alpha-helical content of the protein at 40-50 degrees C, an inactivating effect of charged surfactants and organic co-solvent on the peroxidase activity of ferritin indicate a very important role of the apoprotein in peroxidase function. A possible mechanism of apoferritin participation in peroxidase catalysis is discussed.
Arapova
G S
GS
Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Minsk, Belarus.
Eryomin
A N
AN
Metelitza
D I
DI
D000818:Animals; D001059:Apoproteins / Q000378:metabolism*; D002384:Catalysis; D002942:Circular Dichroism; D003902:Detergents; D004126:Dimethylformamide / Q000737:chemistry; D005293:Ferritins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D006358:Hot Temperature; D007700:Kinetics; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism; D013816:Thermodynamics
D001059:Apoproteins; D003902:Detergents; D004126:Dimethylformamide; D005293:Ferritins; D010544:Peroxidases