title: Structural properties of apocytochrome b5: presence of a stable native core.
abstract: Upon removal of the heme group, the water-soluble fragment of cytochrome b5 adopts a conformation less stable and compact than that of the holoprotein [Huntley, T. E., & Strittmatter, P. (1972) J. Biol. Chem. 247, 4641-4647]. This conformation, imposed by the amino acid sequence alone, has not been described in detail. One- and two-dimensional proton nuclear magnetic resonance spectroscopy techniques were applied to the apoprotein of the soluble fragment of rat liver cytochrome b5 in an effort to characterize the structure of the apoprotein. Nuclear Overhauser spectroscopy revealed a number of short interresidue distances and demonstrated that, in spite of the increased flexibility, at least one cluster of side chains exists on a time scale long enough for study. Several residues participating in the cluster, in particular the only Trp (Trp 22), were identified. Similarities with the spectrum of the reduced holoprotein were observed that led to the inspection of the cytochrome b5 crystal structure for assigning resonances. It appeared that the environment of this residue maintains its integrity in the apoprotein. Since in the holoprotein Trp 22 belongs to a hydrophobic core formed in part by beta-strands, it is proposed that some of this beta-structure is stable in the absence of the heme-protein interactions. Implications for structure and folding are discussed.
authors.affiliation: Department of Chemistry, Pennsylvania State University, University Park 16802.
authors.forename: C D
authors.initials: CD
authors.lastname: Moore
authors.forename: J T
authors.initials: JT
authors.lastname: Lecomte
mesh_terms: D000818:Animals; D001059:Apoproteins; D003573:Cytochrome b Group; D045303:Cytochromes b; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011487:Protein Conformation; D051381:Rats; D013696:Temperature; D014867:Water
chemical_list: D001059:Apoproteins; D003573:Cytochrome b Group; D014867:Water; D045303:Cytochromes b