title: Kinetics of the reconstitution of hemoglobin from semihemoglobins alpha and beta with heme.
abstract: Kinetics of the reconstitution of hemoglobin from semihemoglobins alpha and beta with hemin dicyanide have been investigated using three kinds of stopped-flow technique (Soret absorption, fluorescence quenching of tryptophan, and Soret CD). The semihemoglobins alpha and beta are occupied by heme in the alpha and beta chains, respectively, the other chain being heme-free. Based on the kinetic results, the following scheme for the reconstitution is proposed; First, hemin dicyanide enters the pocket-like site of the apo chains. Second, in semihemoglobin alpha, the CN-ligand in the fifth coordination position of iron is replaced by the imidazole ring of the proximal His immediately after the heme insertion. In contrast, semihemoglobin beta changes its conformation after the heme insertion, and this is followed by the ligand replacement. Finally, the partial structure changes induced by the ligand replacement propagate onto the whole molecule and the final conformation is attained. The results indicate that semihemoglobin alpha retains a more rigid and organized structure, and more closely approaches its final structure than does semihemoglobin beta.
authors.affiliation: Department of Biophysical Chemistry, Kitasato University, School of Medicine, Kanagawa, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawamura-Konishi
authors.forename: K
authors.initials: K
authors.lastname: Chiba
authors.forename: H
authors.initials: H
authors.lastname: Kihara
authors.forename: H
authors.initials: H
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D002942:Circular Dichroism; D006418:Heme / Q000737:chemistry; D006427:Hemin / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry*; D007700:Kinetics; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
chemical_list: D001059:Apoproteins; D006454:Hemoglobins; C023478:apohemoglobin; D006418:Heme; D006427:Hemin