Study of the specific heme orientation in reconstituted hemoglobins.
NMR studies of the recombination reaction of apohemoglobin derivatives with natural and unnatural hemes and of the heme-exchange reaction for reconstituted hemoglobin have revealed that the heme is incorporated into the apoprotein with stereospecific heme orientations dependent upon the heme peripheral 2,4-substituents and the axial iron ligand(s). Heme orientations also depend on whether recombination occurs at the alpha or beta subunit and on whether or not the complementary subunit is occupied by the heme. In the recombination reaction with the azido complex of deuterohemin, the alpha subunit of the apohemoglobin preferentially combines with the hemin in the "disordered" heme orientation, whereas protohemin is inserted in either of two heme orientations. Mesohemin inserts predominantly in the "native" heme orientation. For the beta subunit, specific heme orientation was also encountered, but the specificity was somewhat different from that of the alpha subunit. It was also shown that the specific heme orientation in both subunits is substantially affected by the axial heme ligands. These findings imply that apohemoglobin senses the steric bulkiness of both the porphyrin 2,4-substituents and the axial iron ligands in the heme-apoprotein recombination reaction. To gain an insight into the effect of the protein structure, the heme reconstitution reaction of semihemoglobin, demonstrating that the heme orientation in the reconstituted semihemoglobin with the azido-deuterohemin complex was in the native form, was also examined.(ABSTRACT TRUNCATED AT 250 WORDS)
Ishimori
K
K
Division of Molecular Engineering, Graduate School of Engineering, Kyoto University, Japan.
Morishima
I
I
D000328:Adult; D001665:Binding Sites; D006418:Heme; D006441:Hemoglobin A; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D011487:Protein Conformation
D008024:Ligands; D006418:Heme; D006441:Hemoglobin A