The heme iron VDOS reveals that the presence of a second
metal in the nonheme site influences NO binding to FeII-FeBMb1(57FeII) and ZnII-FeBMb1(57FeII). Fe–NO stretching
vibrations, clearly resolved above 400 cm–1, probe
the axial ligation. For reference, dashed lines indicate Fe–NO
stretching frequencies reported for native horse heart MbNO (452 cm–1),93 characteristic of
a six-coordinate complex with NO coordinated trans to a histidine
ligand, and for Fe(DPIX)(NO) (528 cm–1),88 a typical five-coordinate heme NO complex. A
substantial fraction of hemes exhibit an Fe–NO stretching frequency
characteristic of five-coordinate heme nitrosyls when either ZnII or FeII is present in the nonheme site. This
contrasts with previous measurements on native MbNO,92 which revealed a NRVS signal consistent with six-coordinate
heme NO.