title: House fly cytochrome b5 exhibits kinetically trapped hemin and selectivity in hemin binding.
abstract: We report that cytochrome b(5) (cyt b(5)) from Musca domestica (house fly) is more thermally stable than all other microsomal (Mc) cytochromes b(5) that have been examined to date. It also exhibits a much higher barrier to equilibration of the two isomeric forms of the protein, which differ by a 180 degrees rotation about the alpha-gamma-meso axis of hemin (ferric heme). In fact, hemin is kinetically trapped in a nearly statistical 1.2:1 ratio of rotational forms in freshly expressed protein. The equilibrium ratio (5.5:1) is established only upon incubation at temperatures above 37 degrees C. House fly Mc cyt b(5) is only the second b-hemoprotein that has been shown to exhibit kinetically trapped hemin at room temperature or above, the first being cyt b(5) from the outer membrane of rat liver mitochondria (rat OM cyt b(5)). Finally, we show that the small excess of one orientational isomer over the other in freshly expressed protein results from selective binding of hemin by the apoprotein, a phenomenon that has not heretofore been established for any apocyt b(5).
authors.affiliation: Department of Chemistry, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: Lijun
authors.initials: L
authors.lastname: Wang
authors.forename: Ramona J
authors.initials: RJ
authors.lastname: Bieber Urbauer
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Urbauer
authors.forename: David R
authors.initials: DR
authors.lastname: Benson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D015786:Cytochromes b5 / Q000737:chemistry / Q000378:metabolism*; D006427:Hemin / Q000378:metabolism*; D006358:Hot Temperature; D006793:Houseflies; D007700:Kinetics; D024101:Mitochondrial Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D051381:Rats; D016415:Sequence Alignment
chemical_list: D001059:Apoproteins; D024101:Mitochondrial Proteins; D006427:Hemin; D015786:Cytochromes b5