Pulse radiolysis studies of the reactions of CO3•- and NO2• with nitrosyl(II)myoglobin and nitrosyl(II)hemoglobin

The reactions of carbonate radical anion [CO3 center dot-, systematic name: trioxidocarbonate(center dot 1-)] with nitrosyl(II)hemoglobin (HbFe(II)NO) and nitrosyl(II)myoglobin (MbFe(II)NO) were studied by pulse radiolysis in N2O-saturated 0.25 M sodium bicarbonate solutions at pH 10.0 and room temperature. The reactions proceed in two steps: outer-sphere oxidation of the nitrosyliron(II) proteins to their corresponding nitrosyliron(III) forms and subsequent dissociation of NO center dot. The second-order rate constants measured for the first reaction steps were (4.3 +/- 0.2) x 10(8) and (1.5 +/- 0.3) x 10(8) M-1 s(-1), for MbFe(II)NO and HbFe(II)NO, respectively. The reactions between nitrogen dioxide and MbFe(II)NO or HbFe(II)NO were studied by pulse radiolysis in N2O-saturated 0.1 M phosphate buffer pH 7.4 containing 5 mM nitrite. Also for the reactions of this oxidant with the nitrosyliron(IT) forms of Mb and Hb a two-step reaction was observed: oxidation of the iron was followed by dissociation of NO center dot. The second-order rate constants measured for the first reaction steps were (2.9 +/- 0.3) x 10(7) and (1.8 +/- 0.3) x 10(7) M-1 s(-1), for MbFe(II)NO and HbFe(II)NO, respectively. Both radicals appear to be able to oxidize the iron(II) centers of the proteins directly. Only for the reactions with HbFe(II)NO it cannot be excluded that, in a parallel reaction, CO3 center dot- and NO2 center dot first react with amino acid(s) of the globin, which then oxidize the nitrosyliron(II) center.