title: Enzymatic characterization of four new mutations in the glucose-6 phosphatase (G6PC) gene which cause glycogen storage disease type 1a.
abstract: Glycogen storage disease type 1a (GSD1a) is caused by mutations in the gene of glucose-6 phosphatase (G6PC), encoding the last enzyme of gluconeogenesis and glycogenolysis. To study the effect of mutations previously identified, but not yet enzymatically characterized, in French GSD1a patients, we used an in vitro expression system of the human glucose-6 phosphatase (hGlc6Pase) cDNA. Wild type hGlc6Pase expressed in COS-7 cells exhibited kinetic features comparable to microsomal Glc6Pase from normal human liver and kidney. Four new mutations inducing aminoacid changes in the coding sequence, e.g. W77R, A124T, G184E and L211P, were inserted into the Glc6Pase cDNA by site-directed mutagenesis, and studied after transient expression in COS-7 cells. All four mutations totally abolished Glc6Pase activity.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (Unité 449), Faculté de Médecine R. T. H. Laennec, Lyon, France.
authors.forename: N
authors.initials: N
authors.lastname: Bruni
authors.forename: F
authors.initials: F
authors.lastname: Rajas
authors.forename: S
authors.initials: S
authors.lastname: Montano
authors.forename: F
authors.initials: F
authors.lastname: Chevalier-Porst
authors.forename: I
authors.initials: I
authors.lastname: Maire
authors.forename: G
authors.initials: G
authors.lastname: Mithieux
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D015152:Blotting, Northern; D019556:COS Cells; D015971:Gene Expression Regulation, Enzymologic; D005952:Glucose-6-Phosphatase / Q000235:genetics / Q000378:metabolism*; D005953:Glycogen Storage Disease Type I / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D005952:Glucose-6-Phosphatase