title: Solution structure of a conserved domain of antizyme: a protein regulator of polyamines.
abstract: Antizyme and its isoforms are members of an unusual yet broadly conserved family of proteins, with roles in regulating polyamine levels within cells. Antizyme has the ability to bind and inhibit the enzyme ornithine decarboxylase (ODC), targeting it for degradation at the proteasome; antizyme is also known to affect the transport of polyamines and interact with the antizyme inhibitor protein (AZI), as well as the cell-cycle protein cyclin D1. In the present work, NMR methods were used to determine the solution structure of a stable, folded domain of mammalian antizyme isoform-1 (AZ-1), consisting of amino acid residues 87-227. The protein was found to contain eight beta strands and two alpha helices, with the strands forming a mixed parallel and antiparallel beta sheet. At the level of primary sequence, antizyme is not similar to any protein of known structure, and results show that antizyme exhibits a novel arrangement of its strands and helices. Interestingly, however, the fold of antizyme is similar to that found in a family of acetyl transferases, as well as translation initiation factor IF3, despite a lack of functional relatedness between these proteins. Structural results, combined with amino acid sequence comparisons, were used to identify conserved features among the various homologues of antizyme and their isoforms. Conserved surface residues, including a cluster of acidic amino acids, were found to be located on a single face of antizyme, suggesting this surface is a possible site of interaction with target proteins such as ODC. This structural model provides an essential framework for an improved future understanding of how the different parts of antizyme play their roles in polyamine regulation.
authors.affiliation: Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas 78712, USA. dhoffman@mail.utexas.edu
authors.forename: David W
authors.initials: DW
authors.lastname: Hoffman
authors.forename: Donald
authors.initials: D
authors.lastname: Carroll
authors.forename: Nadia
authors.initials: N
authors.lastname: Martinez
authors.forename: Marvin L
authors.initials: ML
authors.lastname: Hackert
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry; D017124:Conserved Sequence; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D065108:Ornithine Decarboxylase Inhibitors; D010446:Peptide Fragments / Q000737:chemistry; D011073:Polyamines / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D051381:Rats; D016415:Sequence Alignment; D012995:Solubility
chemical_list: D002352:Carrier Proteins; D065108:Ornithine Decarboxylase Inhibitors; D010446:Peptide Fragments; D011073:Polyamines; D020033:Protein Isoforms; D011506:Proteins; C031017:ornithine decarboxylase antizyme; C039317:ornithine decarboxylase antizyme inhibitor