heme environment in barley hemoglobin

• Published in: The Journal of biological chemistry 1999-02, Vol.274 (7), p.4207-4212
• Main Authors: Das, T.K, Lee, H.C, Duff, S.M.G, Hill, R.D, Peisach, J, Rousseau, D.L, Wittenberg, B.A, Wittenberg, J.B
• Format: Article
• Language: English
• Subjects: electron paramagnetic resonance spectroscopy; Electron Spin Resonance Spectroscopy; ferric ions; ferrous ions; heme; Heme - chemistry; Hemeproteins - chemistry; hemoglobin; histidine; Histidine - chemistry; Hordeum vulgare; Hydrogen Bonding; Hydrogen-Ion Concentration; iron; Ligands; Plant Proteins - chemistry; Raman spectroscopy; spectral analysis; spectroscopy; Spectrum Analysis, Raman
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.7.4207

To elucidate the environment and ligand structure of the heme in barley hemoglobin (Hb), resonance Raman and electron paramagnetic resonance spectroscopic studies have been carried out. The heme is shown to have bis -imidazole coordination, and neither of the histidines has imidazolate character. Barley Hb has a unique heme environment as judged from the Fe-CO and C-O stretching frequencies in the CO complex. Two Fe-CO stretching modes are observed with frequencies at 534 and 493 cm −1 , with relative intensities that are pH sensitive. The 534 cm −1 conformer shows a deuterium shift, indicating that the iron-bound CO is hydrogen-bonded, presumably to the distal histidine. A C-O stretching mode at 1924 cm −1 is assigned as being associated with the 534 cm −1 conformer. Evidence is presented that the high Fe-CO and low C-O stretching frequencies (534 and 1924 cm −1 , respectively) arise from a short hydrogen bond between the distal histidine and the CO. The 493 cm −1 conformer arises from an open conformation of the heme pocket and becomes the dominant population under acidic conditions when the distal histidine moves away from the CO. Strong hydrogen bonding between the bound ligand and the distal histidine in the CO complex of barley Hb implies that a similar structure may occur in the oxy derivative, imparting a high stability to the bound oxygen. This stabilization is confirmed by the dramatic decrease in the oxygen dissociation rate compared with sperm whale myoglobin.