title: Molecular and enzymatic characterization of a unique carnitine palmitoyltransferase 1A mutation in the Hutterite community.
abstract: Hepatic carnitine palmitoyltransferase 1 (CPT1A) deficiency is a rare disorder of mitochondrial fatty acid oxidation inherited as an autosomal recessive trait. Symptomatology comprises attacks of hypoketotic hypoglycemia with risk of sudden death or neurological sequelae. Only one CPT1A mutation has been reported so far. Identification of the disease-causing mutations allows both insights into the structure-function relationships of CPT1A and management of the patients and their relatives. The molecular analysis of CPT1A deficiency in a large Hutterite kindred illustrates this point. Both cDNA and genomic DNA analysis demonstrate that the affected patients are homozygous for a 2129G>A mutation predicting a G710E substitution. Studies in fibroblasts from one patient as well as heterologous expression of the mutagenized CPT1A in yeast show that the G710E mutation alters neither mitochondrial targeting nor stability of the CPT1A protein. By contrast, kinetic studies conclusively establish that the mutant CPT1A is totally inactive, indicating that the G710E mutation dramatically impairs the catalytic function of CPT1A. Finally, due to a strongly suspected founder effect for the origin of CPT1A deficiency in this Hutterite kindred, identification of this disease-causing mutation allows the setup of a targeted DNA-based newborn screening in this at-risk population.
authors.affiliation: CNRS UPR 1524, Meudon, France.
authors.forename: C
authors.initials: C
authors.lastname: Prip-Buus
authors.forename: L
authors.initials: L
authors.lastname: Thuillier
authors.forename: N
authors.initials: N
authors.lastname: Abadi
authors.forename: C
authors.initials: C
authors.lastname: Prasad
authors.forename: L
authors.initials: L
authors.lastname: Dilling
authors.forename: J
authors.initials: J
authors.lastname: Klasing
authors.forename: F
authors.initials: F
authors.lastname: Demaugre
authors.forename: C R
authors.initials: CR
authors.lastname: Greenberg
authors.forename: J C
authors.initials: JC
authors.lastname: Haworth
authors.forename: V
authors.initials: V
authors.lastname: Droin
authors.forename: N
authors.initials: N
authors.lastname: Kadhom
authors.forename: S
authors.initials: S
authors.lastname: Gobin
authors.forename: P
authors.initials: P
authors.lastname: Kamoun
authors.forename: J
authors.initials: J
authors.lastname: Girard
authors.forename: J P
authors.initials: JP
authors.lastname: Bonnefont
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002334:Carnitine O-Palmitoyltransferase / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005006:Ethnicity / Q000235:genetics*; D005192:Family Health; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D017354:Point Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D018076:DNA, Complementary; D002334:Carnitine O-Palmitoyltransferase