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UV-vis spectral characterization of the reduction of ferric H64A sw MbII–H2O by dithionite, followed by the reaction of ferrous H64A sw deoxyMbII with the nitroalkane precursors to form the respective ferrous H64A sw MbII–RNO adducts. A) H64A sw MbII–MeNO, B) H64A sw MbII–EtNO, C) H64A sw MbII–PrNO, and D) H64A sw MbII–iPrNO. Final reaction conditions: 3 μM H64A sw Mb, 0.1 M phosphate buffer (pH 7.4), 20 mM dithionite, 20 mM RNO2.
