ABCMdb

PMID: 21476988

Zhang X, De Marcos Lousa C, Schutte-Lensink N, Ofman R, Wanders RJ, Baldwin SA, Baker A, Kemp S, Theodoulou FL
Conservation of targeting but divergence in function and quality control of peroxisomal ABC transporters: an analysis using cross-kingdom expression.
Biochem J. 2011 Jun 15;436(3):547-57., [PubMed]

Sentences

No. Mutations Sentence Comment

153 ABCD1 p.Arg660Trp ABCD1 p.Ser149Asn ABCD1 p.Arg554His ABCD1 p.Glu609Lys ABCD1 p.His667Asp ABCD1 p.Leu654Pro ABCD1 p.Ser606Leu ABCD1 p.Arg389His ABCD1 p.Arg104Cys ABCD1 p.Leu220Pro ABCD1 p.Glu609Gly ABCD1 p.Arg74Trp ABCD1 p.Ala616Thr ABCD1 p.Asp194His Approximately 60% of X-ALD ABCD1 mutations are missense mutations, 65% of which result in no detectable ALDP, based on IF (immunofluorescence), indicating that they affect protein Table 1 Quantification of ALDP levels in X-ALD fibroblasts ALDP Mutation IF Immunoblot (% of control) p.Arg74Trp Absent 7.5 + - 0.6 p.Arg104Cys Reduced 35 + - 3.0 p.Ser149Asn Present 77 + - 3.0 p.Asp194His Present 60 + - 13.6 p.Leu220Pro Reduced 21.8 + - 5.4 p.Arg389His Present 40.6 + - 3.6 p.Arg554His Absent 1.0 + - 0.5 p.Ser606Leu Present 25 + - 1.5 p.Glu609Gly Absent 2.1 + - 1.3 p.Glu609Lys Absent 1.8 + - 0.9 p.Ala616Thr Absent 4.3 + - 1.7 p.Leu654Pro Absent 1.5 + - 1.3 p.Arg660Trp Absent 1.6 + - 0.8 p.His667Asp Absent 2.9 + - 1.0 p.Arg113fs Absent - Figure 3 Interaction of mammalian ABCD proteins with Arabidopsis Pex19 in vivo Tobacco plants stably expressing CFP-SKL were co-transfected with 35S::ABCD-YFP fusions andNLS-Pex19constructs.Leafepidermalcellswereimagedusingconfocalmicroscopy:(A-D) ALDP-YFP plus NLS-HsPex19; (E-H) ALDP-YFP plus NLS-AtPex19_1; (I-L) ALDR-YFP plus NLS-AtPex19_1. Login to comment 154 ABCD1 p.Arg660Trp ABCD1 p.Ser149Asn ABCD1 p.Arg554His ABCD1 p.Glu609Lys ABCD1 p.His667Asp ABCD1 p.Leu654Pro ABCD1 p.Ser606Leu ABCD1 p.Arg389His ABCD1 p.Arg104Cys ABCD1 p.Leu220Pro ABCD1 p.Glu609Gly ABCD1 p.Arg74Trp ABCD1 p.Ala616Thr ABCD1 p.Asp194His Approximately 60% of X-ALD ABCD1 mutations are missense mutations, 65% of which result in no detectable ALDP, based on IF (immunofluorescence), indicating that they affect protein Table 1 Quantification of ALDP levels in X-ALD fibroblasts ALDP Mutation IF Immunoblot (% of control) p.Arg74Trp Absent 7.5 + - 0.6 p.Arg104Cys Reduced 35 + - 3.0 p.Ser149Asn Present 77 + - 3.0 p.Asp194His Present 60 + - 13.6 p.Leu220Pro Reduced 21.8 + - 5.4 p.Arg389His Present 40.6 + - 3.6 p.Arg554His Absent 1.0 + - 0.5 p.Ser606Leu Present 25 + - 1.5 p.Glu609Gly Absent 2.1 + - 1.3 p.Glu609Lys Absent 1.8 + - 0.9 p.Ala616Thr Absent 4.3 + - 1.7 p.Leu654Pro Absent 1.5 + - 1.3 p.Arg660Trp Absent 1.6 + - 0.8 p.His667Asp Absent 2.9 + - 1.0 p.Arg113fs Absent - Figure 3 Interaction of mammalian ABCD proteins with Arabidopsis Pex19 in vivo Tobacco plants stably expressing CFP-SKL were co-transfected with 35S::ABCD-YFP fusions andNLS-Pex19constructs.Leafepidermalcellswereimagedusingconfocalmicroscopy:(A-D) ALDP-YFP plus NLS-HsPex19; (E-H) ALDP-YFP plus NLS-AtPex19_1; (I-L) ALDR-YFP plus NLS-AtPex19_1. Login to comment 169 ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Leu654Pro ABCD1 p.Arg104Cys ABCD1 p.Glu609Gly ABCD1 p.Ala616Thr ABCD1 p.Arg74Cys Results are means + - S.D.; n = 3. significantly in response to low temperature in ten wild-type control lines tested (see Supplementary Figure S3A at http://www.BiochemJ.org/bj/436/bj4360547add.htm); however, increased expression levels of ALDP were found in several of the X-ALD fibroblasts investigated: p.Arg74Cys, p.Arg104Cys, p.Arg554His, p.Glu609Gly, p.Ala616Thr, p.Leu654Pro and p.Arg660Trp (Figures 4A and 4B). Login to comment 170 ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Leu654Pro ABCD1 p.Arg104Cys ABCD1 p.Glu609Gly ABCD1 p.Ala616Thr ABCD1 p.Arg74Cys Results are means + - S.D.; n = 3. significantly in response to low temperature in ten wild-type control lines tested (see Supplementary Figure S3A at http://www.BiochemJ.org/bj/436/bj4360547add.htm); however, increased expression levels of ALDP were found in several of the X-ALD fibroblasts investigated: p.Arg74Cys, p.Arg104Cys, p.Arg554His, p.Glu609Gly, p.Ala616Thr, p.Leu654Pro and p.Arg660Trp (Figures 4A and 4B). Login to comment 172 ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Glu609Lys ABCD1 p.Glu609Gly ABCD1 p.Ala616Thr ABCD1 p.Asp194His ALDP was increased from 2-4% to ~20% of wild-type levels in cell lines bearing the mutations p.Glu609Gly, p.Ala616Thr and p.Arg660Trp, from 1 to 10% in p.Glu609Lys and p.Arg554His cells and from 45 to 75% in the p.Asp194His cell line (Figure 4A). Login to comment 173 ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Glu609Lys ABCD1 p.Glu609Gly ABCD1 p.Ala616Thr ABCD1 p.Ala616Thr ABCD1 p.Ala616Thr ABCD1 p.Asp194His VLCFA β-oxidation was measured in cells that were cultured at 30◦ C for 72 h, but in only one case (p.Ala616Thr) was the capacity to degrade VLCFA restored to near-control levels (Figure 4C). Login to comment 174 ABCD1 p.Arg660Trp ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Arg554His ABCD1 p.Ala616Thr ABCD1 p.Ala616Thr ABCD1 p.Ala616Thr However, after 4 weeks of culture at 30◦ C, VLCFA levels were partially corrected in p.Arg660Trp, p.Arg554His and p.Ala616Thr fibroblasts. Login to comment 175 ABCD1 p.Arg660Trp ABCD1 p.Arg554His ABCD1 p.Ala616Thr However, after 4 weeks of culture at 30e6; C, VLCFA levels were partially corrected in p.Arg660Trp, p.Arg554His and p.Ala616Thr fibroblasts. Login to comment 176 ABCD1 p.Glu609Gly ABCD1 p.Asp194His Lines bearing the p.Glu609Gly and p.Asp194His mutations did not show any correction of the C26:0/C22:0 ratio (Figure 4D). Login to comment 177 ABCD1 p.Glu609Gly ABCD1 p.Asp194His Lines bearing the p.Glu609Gly and p.Asp194His mutations did not show any correction of the C26:0/C22:0 ratio (Figure 4D). Login to comment 181 ABCD1 p.Tyr174Cys ABCD1 p.His667Asp ABCD1 p.Arg617His ABCD1 p.Ser606Leu ABCD1 p.Arg104Cys These included relatively common mutants which are unstable in fibroblasts, but which can be rescued by the application of proteasome inhibitors (p.Ser606Leu, p.Arg617His and p.His667Asp), the p.Arg104Cys mutant in which degradation of ALDP cannot be prevented by proteasome inhibitors and the p.Tyr174Cys mutant which is Table 2 X-ALD mutants used for analysis of targeting in tobacco cells The occurrence is the number of documented patients bearing the mutation; source: X-ALD database (http://www.x-ald.nl). Login to comment 182 ABCD1 p.Tyr174Cys ABCD1 p.His667Asp ABCD1 p.Arg617His ABCD1 p.Ser606Leu ABCD1 p.Arg104Cys These included relatively common mutants which are unstable in fibroblasts, but which can be rescued by the application of proteasome inhibitors (p.Ser606Leu, p.Arg617His and p.His667Asp), the p.Arg104Cys mutant in which degradation of ALDP cannot be prevented by proteasome inhibitors and the p.Tyr174Cys mutant which is c The Authors Journal compilation c Table 2 X-ALD mutants used for analysis of targeting in tobacco cells The occurrence is the number of documented patients bearing the mutation; source: X-ALD database (http://www.x-ald.nl). Login to comment