title: Purification and characterization of aspartate N-acetyltransferase: A critical enzyme in brain metabolism.
abstract: Canavan disease (CD) is a neurological disorder caused by an interruption in the metabolism of N-acetylaspartate (NAA). Numerous mutations have been found in the enzyme that hydrolyzes NAA, and the catalytic activity of aspartoacylase is significantly impaired in CD patients. Recent studies have also supported an important role in CD for the enzyme that catalyzes the synthesis of NAA in the brain. However, previous attempts to study this enzyme had not succeeded in obtaining a soluble, stable and active form of this membrane-associated protein. We have now utilized fusion constructs with solubilizing protein partners to obtain an active and soluble form of aspartate N-acetyltransferase. Characterization of the properties of this enzyme has set the stage for the development of selective inhibitors that can lower the elevated levels of NAA that are observed in CD patients and potentially serve as a new treatment therapy.
authors.affiliation: Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606, USA.
authors.forename: Qinzhe
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606, USA.
authors.forename: Mojun
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606, USA.
authors.forename: Gwenn G
authors.initials: GG
authors.lastname: Parungao
authors.affiliation: Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606, USA. Electronic address: ron.viola@utoledo.edu.
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Viola
mesh_terms: D000123:Acetyltransferases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001921:Brain / Q000201:enzymology; D002846:Chromatography, Affinity; D017124:Conserved Sequence; D004795:Enzyme Stability; D004926:Escherichia coli; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011993:Recombinant Fusion Proteins; D000123:Acetyltransferases; C000611572:NAT8L protein, human
keywords: Canavan disease; aspartate N-acetyltransferase; enzyme kinetics; enzyme purification; membrane protein