Protein conformation change of myoglobin upon ligand binding probed by ultraviolet resonance Raman spectroscopy.
Conformational change of myoglobin (Mb) accompanied by binding of a ligand was investigated with 244 nm excited ultraviolet resonance Raman Spectroscopy (UVRR). The UVRR spectra of native sperm whale (sw) and horse (h) Mbs and W7F and W14F swMb mutants for the deoxy and CO-bound states enabled us to reveal the UVRR spectra of Trp7, Trp14, and Tyr151 residues, separately. The difference spectra between the deoxy and CO-bound states reflected the environmental or structural changes of Trp and Tyr residues upon CO binding. The W3 band of Trp7 near the N-terminus exhibited a change upon CO binding, while Trp14 did not. Tyr151 in the C-terminus also exhibited a definite change upon CO binding, but Tyr103 and Tyr146 did not. The spectral change of Tyr residues was characterized through solvent effects of a model compound. The corresponding spectral differences between CO- and n-butyl isocyanide-bound forms were much smaller than those between the deoxy and CO-bound forms, suggesting that the conformation change in the C- and N-terminal regions is induced by the proximal side of the heme through the movement of iron. Although the swinging up of His64 upon binding of a bulky ligand is noted by X-ray crystallographic analysis, UVRR spectra of His for the n-butyl isocyanide-bound form did not detect the exposure of His64 to solvent.
Haruta
N
N
School of Mathematical and Physical Sciences, The Graduate University for Advanced Studies, and Institute for Molecular Science and Center for Integrative Bioscience, Okazaki National Research Institutes, Myodaiji, Okazaki, 444-8585 Japan.
Aki
M
M
Ozaki
S
S
Watanabe
Y
Y
Kitagawa
T
T
D000818:Animals; D017666:Deuterium Oxide / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006736:Horses; D006860:Hydrogen Bonding; D007834:Lasers; D008024:Ligands; D009211:Myoglobin / Q000031:analogs & derivatives* / Q000737:chemistry*; D011487:Protein Conformation; D011524:Protoporphyrins / Q000737:chemistry; D012996:Solutions; D013059:Spectrum Analysis, Raman / Q000379:methods*; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry; D014466:Ultraviolet Rays; D014907:Whales
D008024:Ligands; D009211:Myoglobin; D011524:Protoporphyrins; D012996:Solutions; C018013:carboxymyoglobin; C055547:deoxymyoglobin; D014443:Tyrosine; D006639:Histidine; D014364:Tryptophan; D017666:Deuterium Oxide