title: Simultaneous purification and characterization of aspartate aminotransferase isoenzymes from chicken liver.
abstract: Cytosolic and mitochondrial isoenzymes of aspartate aminotransferase (EC 2.6.1.1) were purified to homogeneity from chicken liver, without previous fractionation of the subcellular components. The procedure includes initial heat treatment and ammonium sulfate fractionation. The two isoenzymes can then be separated by a DEAE-Sepharose chromatography using a linear gradient of L-aspartate (reaction substrate). The separated fractions can be further purified by a parallel step with HA-Ultrogel prior to octyl-Sepharose (c-AAT) and CM-Sepharose (m-AAT) chromatographies. Michaelis constants, pI values, inhibition by adipate and subforms generation with time were studied for both isoenzymes.
authors.affiliation: Departament de Bioquímica i Fisiologia, Facultat de Química, Universitat de Barcelona, Spain.
authors.forename: R
authors.initials: R
authors.lastname: Gracia
authors.forename: M
authors.initials: M
authors.lastname: Busquets
authors.forename: M
authors.initials: M
authors.lastname: Gil
authors.forename: A
authors.initials: A
authors.lastname: Cortés
authors.forename: J
authors.initials: J
authors.lastname: Bozal
mesh_terms: D000818:Animals; D001219:Aspartate Aminotransferases / Q000302:isolation & purification* / Q000378:metabolism; D002645:Chickens; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*
chemical_list: D007527:Isoenzymes; D001219:Aspartate Aminotransferases