Single-crystal EPR study of novel azide complex of cyanogen bromide-modified myoglobin. Influence of specific modification of the heme distal histidyl residue on the ligand-binding structure.
Stable azide complex of cyanogen bromide-modified met-myoglobin (metMb) was prepared and crystallized. The principal values and eigen vectors of g-tensor were determined by single-crystal EPR spectroscopy at 77 K: gxx = 1.50, gyy = 2.32, and gzz = 2.91. These g values were similar to those of tetrazole derivative rather than azide derivative of native metMbs, suggesting that tetrazole derivative might be formed from N-cyanoimidazole of distal histidyl residue via nucleophilic attack of azide ion by 1,3-dipolar cycloaddition reaction. The orientation of the maximal g value (gzz) of the novel product was found to deviate about 13 degrees from the heme normal of native aquometMb. Thus, the orientation of the heme plane might be altered in passing from native metMb to cyanogen bromide-mediated metmyoglobin. The present EPR results demonstrated that the modification of the histidyl residue at the heme distal side causes the changes in the stereochemical and electronic natures of the ligand binding to the heme.
Hori
H
H
Department of Biophysical Engineering, Faculty of Engineering Science, Osaka University, Japan.
Fujii
M
M
Shiro
Y
Y
Iizuka
T
T
Adachi
S
S
Morishima
I
I
D001386:Azides; D001665:Binding Sites; D003488:Cyanogen Bromide / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D006639:Histidine; D008024:Ligands; D009211:Myoglobin / Q000378:metabolism*; D011487:Protein Conformation
D001386:Azides; D008024:Ligands; D009211:Myoglobin; D006639:Histidine; D003488:Cyanogen Bromide