title: Site-directed mutagenesis of bacterial hemoglobin: the role of glutamine (E7) in oxygen-binding in the distal heme pocket.
abstract: The bacterial and yeast hemoglobins have a glutamine instead of histidine in the E7 position of the distal heme pocket. The recently determined crystal structure of Vitreoscilla hemoglobin (VHb) indicates that this residue is oriented out of the heme pocket and may not ligand the bound oxygen. This is in contrast to elephant myoglobin which also has a Gln(E7) but which does ligand the bound oxygen. This residue was changed in VHb using site-directed mutagenesis to leucine (VHbL) or to histidine (VHbH). Spectral and kinetic studies of the binding of oxygen and CO to VHbL showed that this substitution had little effect on the ligand-binding properties of this protein, evidence that Gln(E7) does not H-bond the bound ligand, in agreement with the findings of the crystallographic study of VHb. In contrast, the functional properties of VHbH were drastically altered in a way suggesting that the E7His may itself be liganded to the heme iron. These studies are further evidence that the distal heme pocket in VHb and related microbial hemoglobins differs from that in mammalian hemoglobins and may resemble in some ways the heme pocket in cytochrome b5.
authors.affiliation: Institute of Microbial Technology, Chandigarh, India.
authors.forename: K L
authors.initials: KL
authors.lastname: Dikshit
authors.forename: Y
authors.initials: Y
authors.lastname: Orii
authors.forename: N
authors.initials: N
authors.lastname: Navani
authors.forename: S
authors.initials: S
authors.lastname: Patel
authors.forename: H Y
authors.initials: HY
authors.lastname: Huang
authors.forename: B C
authors.initials: BC
authors.lastname: Stark
authors.forename: D A
authors.initials: DA
authors.lastname: Webster
mesh_terms: D001419:Bacteria / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D005973:Glutamine / Q000378:metabolism; D006454:Hemoglobins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism
chemical_list: D006454:Hemoglobins; D005973:Glutamine; D010100:Oxygen