title: Mutational and protein analysis of patients and heterozygous women with X-linked adrenoleukodystrophy.
abstract: X-linked adrenoleukodystrophy (ALD), a neurodegenerative disorder associated with impaired beta-oxidation of very-long-chain fatty acids (VLCFA), is due to mutations in a gene encoding a peroxisomal ATP-binding cassette (ABC) transporter (ALD protein [ALDP]). We analyzed the open reading frame of the ALD gene in 44 French ALD kindred by using SSCP or denaturing gradient-gel electrophoresis and studied the effect of mutations on ALDP by immunocytofluorescence and western blotting of fibroblasts and/or white blood cells. Mutations were detected in 37 of 44 kindreds and were distributed over the whole protein-coding region, with the exception of the C terminus encoded in exon 10. Except for two mutations (delAG1801 and P560L) observed four times each, nearly every ALD family has a different mutation. Twenty-four of 37 mutations were missense mutations leading to amino acid changes located in or close to putative transmembrane segments (TMS 2, 3, 4, and 5), in the EAA-like motif and in the nucleotide fold of the ATP-binding domain of ALDP. Of 38 ALD patients tested, 27 (71%) lacked ALDP immunoreactivity in their fibroblasts and/or white blood cells. More than half of missense mutations studied (11 of 21) resulted in a complete lack of ALDP immunoreactivity, and six missense mutations resulted in decreased ALDP expression. The fibroblasts and/or white blood cells of 15 of 15 heterozygous carrier from ALD kindred with no ALDP showed a mixture of positive- and negative-ALDP immunoreactivity due to X-inactivation. Since 5%-15% of heterozygous women have normal VLCFA levels, the immunodetection of ALDP in white blood cells can be applicable in a majority of ALD kindred, to identify heterozygous women, particularly when the ALD gene mutation has not yet been identified.
authors.affiliation: INSERM U342, Hôpital Saint-Vincent de Paul, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Feigenbaum
authors.forename: G
authors.initials: G
authors.lastname: Lombard-Platet
authors.forename: S
authors.initials: S
authors.lastname: Guidoux
authors.forename: C O
authors.initials: CO
authors.lastname: Sarde
authors.forename: J L
authors.initials: JL
authors.lastname: Mandel
authors.forename: P
authors.initials: P
authors.lastname: Aubourg
mesh_terms: D000074063:ATP Binding Cassette Transporter, Subfamily D, Member 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000224:Addison Disease / Q000235:genetics; D000293:Adolescent; D000326:Adrenoleukodystrophy / Q000235:genetics*; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004247:DNA / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D017931:DNA Primers; D004586:Electrophoresis; D005091:Exons; D005260:Female; D005347:Fibroblasts; D016368:Frameshift Mutation; D006580:Genetic Carrier Screening; D006801:Humans; D007962:Leukocytes; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D014960:X Chromosome
chemical_list: C115572:ABCD1 protein, human; D000074063:ATP Binding Cassette Transporter, Subfamily D, Member 1; D018528:ATP-Binding Cassette Transporters; D017931:DNA Primers; D008565:Membrane Proteins; D004247:DNA