title: A comparative study on conformation and ligand binding of the neuronal uncoupling proteins.
abstract: Mitochondrial uncoupling proteins of the nervous system (UCPs 2, 4, and 5) have potential roles in the function and protection of the central nervous system (CNS). In the absence of structural information, conformations of the hexahistidine-tagged versions of all five human UCPs in liposomes were investigated for the first time, using far- and near-UV CD and fluorescence spectroscopy. Highly pure UCPs 1-5 were reconstituted in detergents and stable small unilamellar vesicles, appropriate for spectroscopic studies. All UCPs formed dominantly helical conformations in negatively charged phospholipid vesicles (palmitoyloleoylphosphatidylcholine/palmitoyloleoylphosphatidylglycerol, 7:3 molar ratio). UCPs 2 and 5 exhibited comparable helical conformations with possible association in lipid bilayers, whereas UCP4 had a different helical profile that can be related to its less associated form. Interaction of reconstituted UCPs with GDP and GTP, inhibitors of the prototypic UCP1, was detected by near-UV CD and fluorescence spectroscopy, utilizing the sensitivity of these techniques to microenvironments around Trp residues close to the nucleotide binding site. Binding of UCP4 to purine nucleotides was also different from other UCPs. Binding of fatty acids, activators of proton transport in UCPs, to UCPs could not be unambiguously detected, implying a nonbinding conformation/orientation of the proteoliposomes. Interaction of CoA with UCPs was comparable to nucleotide binding, suggesting a possible binding of this molecule at the nucleotide binding site. Despite dissimilar primary sequences, neuronal UCPs share common structural and functional properties with UCPs 1 and 3, supporting a common physiological role in addition to their specific roles in the CNS.
authors.affiliation: Departments of Chemistry, Wilfrid Laurier University, Waterloo, Ontario N2L 3C5, Canada.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Ivanova
authors.forename: Tuan
authors.initials: T
authors.lastname: Hoang
authors.forename: Fern R
authors.initials: FR
authors.lastname: McSorley
authors.forename: Gabriela
authors.initials: G
authors.lastname: Krnac
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Smith
authors.forename: Masoud
authors.initials: M
authors.lastname: Jelokhani-Niaraki
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D006801:Humans; D007473:Ion Channels / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000378:metabolism*; D000071245:Mitochondrial Uncoupling Proteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D016415:Sequence Alignment; D013050:Spectrometry, Fluorescence; D000071256:Uncoupling Protein 1; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3
chemical_list: D007473:Ion Channels; D008024:Ligands; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D000071245:Mitochondrial Uncoupling Proteins; D009419:Nerve Tissue Proteins; C116584:SLC25A14 protein, human; C117577:SLC25A27 protein, human; C000604371:UCP1 protein, human; D000071256:Uncoupling Protein 1; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3