title: Very high resolution structure of a trematode hemoglobin displaying a TyrB10-TyrE7 heme distal residue pair and high oxygen affinity.
abstract: Monomeric hemoglobin from the trematode Paramphistomum epiclitum displays very high oxygen affinity (P(50)<0.001 mm Hg) and an unusual heme distal site containing tyrosyl residues at the B10 and E7 positions. The crystal structure of aquo-met P. epiclitum hemoglobin, solved at 1.17 A resolution via multiwavelength anomalous dispersion techniques (R-factor=0.121), shows that the heme distal site pocket residue TyrB10 is engaged in hydrogen bonding to the iron-bound ligand. By contrast, residue TyrE7 is unexpectedly locked next to the CD globin region, in a conformation unsuitable for heme-bound ligand stabilisation. Such structural organization of the E7 distal residue differs strikingly from that observed in the nematode Ascaris suum hemoglobin (bearing TyrB10 and GlnE7 residues), which also displays very high oxygen affinity. The oxygenation and carbonylation parameters of wild-type P. epiclitum Hb as well as of single- and double-site mutants, with residue substitutions at positions B10, E7 and E11, have been determined and are discussed here in the light of the protein atomic resolution crystal structure.
authors.affiliation: Department of Physics-INFM, Advanced Biotechnology Centre, University of Genova, Largo Rosanna Benzi, 10, Genova, I-16132, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Pesce
authors.forename: S
authors.initials: S
authors.lastname: Dewilde
authors.forename: L
authors.initials: L
authors.lastname: Kiger
authors.forename: M
authors.initials: M
authors.lastname: Milani
authors.forename: P
authors.initials: P
authors.lastname: Ascenzi
authors.forename: M C
authors.initials: MC
authors.lastname: Marden
authors.forename: M L
authors.initials: ML
authors.lastname: Van Hauwaert
authors.forename: J
authors.initials: J
authors.lastname: Vanfleteren
authors.forename: L
authors.initials: L
authors.lastname: Moens
authors.forename: M
authors.initials: M
authors.lastname: Bolognesi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002248:Carbon Monoxide / Q000378:metabolism; D018360:Crystallography, X-Ray; D006418:Heme / Q000378:metabolism*; D006860:Hydrogen Bonding; D007501:Iron / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008706:Methemoglobin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism*; D010250:Paramphistomatidae / Q000737:chemistry* / Q000235:genetics; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D013816:Thermodynamics; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
chemical_list: D008024:Ligands; D014443:Tyrosine; D006418:Heme; C024963:aquomethemoglobin; D002248:Carbon Monoxide; D008706:Methemoglobin; D007501:Iron; D010100:Oxygen