title: A Novel Double Mutation in the ABCD1 Gene in a Patient with X-linked Adrenoleukodystrophy: Analysis of the Stability and Function of the Mutant ABCD1 Protein.
abstract: We diagnosed an adrenomyeloneuropathy (AMN) patient with a double novel missense mutation, c.284C>A (p.A95D) and c.290A>T (p.H97L) in a single ABCD1 allele. In skin fibroblasts from the patient, no ABCD1 protein was detected by immunoblot analysis, and the C24:0 β-oxidation activity was decreased to a level at which the ABCD1 protein was absent. To determine the responsible gene mutation in the patient, we constructed three kinds of mutated ABCD1 gene expression vectors (c.284C>A, c.290A>T or c.284C>A/c.290A>T) and transfected them into CHO cells stably expressing GFP-SKL (CHO/GFP-SKL cells) or CADDS fibroblasts lacking the ABCD1 gene. ABCD1 (p.H97L) displayed the correct peroxisomal localization in CHO/GFP-SKL cells, but ABCD1 (p.A95D) and ABCD1 (p.A95D/p.H97L) were diffuse in the cytosol. Furthermore, ABCD1 (p.H97L) was detected by immunoblot analysis and restored the C24:0 β-oxidation activity in the CADDS fibroblasts, as the wild type ABCD1 did. On the other hand, ABCD1 (p.A95D) and ABCD1 (p.A95D/p.H97L) were not detected and the C24:0 β-oxidation activity was not restored. These results clearly show that c.284C>A is the responsible gene mutation, whereas c.290A>T is a novel polymorphism.
authors: Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, Japan.
authors: Masashi
authors: M
authors: Morita
authors: Junpei
authors: J
authors: Kobayashi
authors: Kozue
authors: K
authors: Yamazaki
authors: Kosuke
authors: K
authors: Kawaguchi
authors: Ayako
authors: A
authors: Honda
authors: Kenji
authors: K
authors: Sugai
authors: Nobuyuki
authors: N
authors: Shimozawa
authors: Reiji
authors: R
authors: Koide
authors: Tsuneo
authors: T
authors: Imanaka