ABCMdb

PMID: 16412981

Gueugnon F, Volodina N, Taouil JE, Lopez TE, Gondcaille C, Grand AS, Mooijer PA, Kemp S, Wanders RJ, Savary S
A novel cell model to study the function of the adrenoleukodystrophy-related protein.
Biochem Biophys Res Commun. 2006 Mar 3;341(1):150-7. Epub 2006 Jan 6., [PubMed]

Sentences

No. Mutations Sentence Comment

42 ABCD2 p.Asp207His Site directed mutagenesis was performed on the ALDRP-EGFP-pTRE2hyg to obtain the mutated construct D207H-ALDRP-EGFP-pTRE2hyg (GAC codon 619 switched to CAC). Login to comment 43 ABCD1 p.Asp194His This mutation in ALDRP, located in a region highly conserved between ALDRP and ALDP, corresponds to the D194H mutation in ALDP, a mutation found in X-ALD patients (see www.x-ald.nl), which disrupts the function without alteration of the peroxisomal localization and probably of the dimerization ability [30]. Login to comment 86 ABCD2 p.Asp207His One microgram of the ALDRP, ALDRP-EGFP, or D207H- ALDRP-EGFP pcDNA3.1/Zeo(+) constructs was used as template in a final volume of 50 ll. Login to comment 149 ABCD2 p.Asp207His or with D207H-ALDRP-EGFP-pTRE2hyg. Login to comment 162 ABCD2 p.Asp207His Taken together, the results indicate that the clones 28 and 19 express ALDRP-EGFP and D207H-ALDRP-EGFP, respectively, at a high level in the peroxisome upon doxycycline treatment. Login to comment 165 ABCD2 p.Asp207His Maximum level of induction (9-fold for the ALDRP-EGFP mRNA and 15-fold for D207H-ALDRP-EGFP mRNA) was reached with 2 lg/ml doxycycline. Login to comment 170 ABCD2 p.Asp207His Analysis of the expression and of the subcellular localization of the normal and mutated (D207H) ALDRP-EGFP. Login to comment 171 ABCD2 p.Asp207His (A) In vitro translation of the ALDRP, ALDRP-EGFP, or D207H-ALDRP-EGFP pcDNA3.1/Zeo(+) constructs using L[35 S]methionine. Login to comment 187 ABCD2 p.Asp207His Characterization of the H4IIEC3-TetOn cell clones expressing a normal (clone 28) or mutated (D207H, clone 19) ALDRP-EGFP fusion protein upon doxycycline treatment. Login to comment 199 ABCD2 p.Asp207His Quantification of the relative expression levels of mRNA of the fusion proteins by quantitative real-time RT-PCR with primers against Abcd2 in the H4IIEC3-TetOn cell clones expressing normal ALDRP-EGFP protein-clone 28 (open rectangles) and mutated (D207H) ALDRP-EGFP protein-clone 19 (filled rectangles). Login to comment 214 ABCD2 p.Asp207His ABCD1 p.Asp194His The D207H mutation in ALDRP is similar to the D194H mutation in ALDP that has been previously studied by stable transfection and has been shown to disrupt function without suppressing the capacity of dimerization and of import in the peroxisomal membrane [30]. Login to comment