title: Studies on dilution inactivation of sheep liver pyruvate carboxylase.
abstract: When sheep liver pyruvate carboxylase was diluted below 4 EU/ml, it underwent inactivation involving two kinetically distinct processes, i.e., a rapid initial burst followed by a slower second phase. The catalytic activity of the diluted enzyme eventually approached zero, suggesting the occurrence of an irreversible process. Analysis of the quaternary structure of the enzyme by gel filtration chromatography and electron microscopy showed that most of the enzyme molecules occur as tetramers at high enzyme concentrations. However, dilution of the enzyme below 4 EU/ml led to the appearance of dimers and monomers which were essentially inactive under the conditions of the assay system used. The presence of acetyl-CoA during dilution prevented inactivation from occurring and preserved the tetrameric structure. When added after dilution, acetyl-CoA prevented further inactivation from occurring but did not reactivate the enzyme. However, acetyl-CoA did cause a relatively rapid reassociation of the inactive monomers and dimers to form inactive tetramers.
authors.affiliation: Department of Biochemistry, University of Adelaide, South Australia.
authors.forename: Y S
authors.initials: YS
authors.lastname: Khew-Goodall
authors.forename: W
authors.initials: W
authors.lastname: Johannssen
authors.forename: P V
authors.initials: PV
authors.lastname: Attwood
authors.forename: J C
authors.initials: JC
authors.lastname: Wallace
authors.forename: D B
authors.initials: DB
authors.lastname: Keech
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D011766:Pyruvate Carboxylase / Q000378:metabolism*; D012756:Sheep; D012996:Solutions; D013329:Structure-Activity Relationship
chemical_list: D046911:Macromolecular Substances; D012996:Solutions; D000105:Acetyl Coenzyme A; D011766:Pyruvate Carboxylase