Changes in the abnormal alpha-subunit upon CO-binding to the normal beta-subunit of Hb M Boston: resonance Raman, EPR and CD study.
Heme-heme interaction in Hb M Boston (His alpha 58-->Tyr) was investigated with visible and UV resonance Raman (RR), EPR, and CD spectroscopies. Although Hb M Boston has been believed to be frozen in the T quaternary state, oxygen binding exhibited appreciable co-operativity (n=1.4) and the near-UV CD spectrum indicated weakening of the T marker at pH 9.0. Binding of CO to the normal beta-subunit gave no change in the EPR and visible Raman spectra of the abnormal alpha-subunit at pH 7.5, but it caused an increase of EPR rhombicity and significant changes in the Raman coordination markers as well as the Fe(III)-tyrosine related bands of the alpha-subunit at pH 9.0. The UVRR spectra indicated appreciable changes of Trp but not of Tyr upon CO binding to the alpha-subunit at pH 9.0. Therefore, we conclude that the ligand binding to the beta heme induces quaternary structure change at pH 9.0 and is communicated to the alpha heme, presumably through His beta 92-->Trp beta 37-->His alpha 87.
Center for Integrative Bioscience, Okazaki National Research Institutes, Myodaiji, Okazaki, 444-8585, Aichi, Japan.
Shigenori
S
Nagatomo
Yayoi
Y
Jin
Masako
M
Nagai
Hiroshi
H
Hori
Teizo
T
Kitagawa
D000596:Amino Acids / Q000097:blood / Q000235:genetics; D002248:Carbon Monoxide / Q000097:blood* / Q000737:chemistry; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D006449:Hemoglobin M / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000097:blood / Q000378:metabolism; D010100:Oxygen / Q000097:blood / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary / Q000235:genetics; D013059:Spectrum Analysis, Raman
D000596:Amino Acids; C027541:hemoglobin M Boston; D002248:Carbon Monoxide; D006449:Hemoglobin M; D007501:Iron; D010100:Oxygen