Anomalous pH dependence of the heme-bound carbon monoxide spectroscopic properties in the Glycera dibranchiata monomer hemoglobin fraction compared to vertebrate hemoglobins

Abstract

The pH dependence of infrared and NMR spectroscopic parameters for carbon monoxide bound to human, equine, rabbit and Glycera dibranchiata monomer fraction hemoglobins has been examined. In all cases, the vertebrate hemoglobins exhibit CO vibrations and 13CO chemical shifts which are pH dependent, whereas the invertebrate hemoglobin does not. The Glycera dibranchiata monomer fraction exhibits the highest wavenumber CO vibration (1970 cm-1) and the most shielded chemical shift (206.2 ppm). The pH behavior of the vertebrate CO-hemoglobins is that the heme-coordinated carbon monoxide chemical shifts and principal infrared vibrations tend toward the values observed for the G. dibranchiata CO-hemoglobin fraction. These results are interpreted as originating in protonation of the distal histidine (E-7) in the vertebrate hemoglobins. The anomalous values for Glycera dibranchiata are concluded to be due to the absence of a distal histidine (E-7 His----Leu) in the heme pocket and not to gross structural dissimilarities between the proteins of the different species examined. Primary sequence similarity matrices have been constructed to compare the functional classes of amino acids at homologous positions for the CD and E helices and for the primary heme contacts in human, equine, sperm whale myoglobin, and the Glycera dibranchiata monomer hemoglobin to illustrate this point. They reveal a high correspondence for all globins and do not correlate with the spectroscopic parameters of heme-coordinated CO.