ABCD1 p.Ser606Leu
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PMID: 11748843
[PubMed]
Kemp S et al: "ABCD1 mutations and the X-linked adrenoleukodystrophy mutation database: role in diagnosis and clinical correlations."
No.
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Comment
174
P560S 7 1678C>T n.d. # P560L 7 1679C>T Reduced P560L 7 1679C>T Reduced fs I588 7 1765delC n.d. # R591P 7 1772G>C Absent S606L 8 1817C>T Present E609K 8 1825G>A Absent E609K 8 1825G>A Absent R617C 8 1849C>T Absent R617H 8 1850G>A Absent R617H 8 1850G>A Absent A626T 9 1876G>A Absent A626T 9 1876G>A Absent A626D 9 1877C>A n.d. # E630G 9 1889A>G n.d. # C631Y 9 1892G>A n.d. # T632I 9 1895C>T n.d. # V635M 9 1903G>A n.d. # L654P 9 1961T>C Absent # R660W 9 1978C>T Absent fs L663 9 1988insT n.d. # fs L663 IVS 9 IVS9+1g>a n.d. # fs L663 IVS 9 IVS9-1g>a n.d. # H667D 10 1999C>G Absent # T668I 10 2003C>T Absent # T693M 10 2078C>T Present # exon1-5del 1-5 n.d. # The 47 mutations marked with a # are novel unique mutations reported for the first time in this paper.
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ABCD1 p.Ser606Leu 11748843:174:120
status: NEW280 For two missense mutations located in functional regions of the ATP-binding domain it was demonstrated that the mutations resulted in either decreased ATP-binding capacity (S606L) or reduced ATPase activity (G512S) [Roerig et al., 2001].
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ABCD1 p.Ser606Leu 11748843:280:173
status: NEW
PMID: 16996397
[PubMed]
Chiu HC et al: "Mutational analyses of Taiwanese kindred with X-linked adrenoleukodystrophy."
No.
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Comment
126
However, common missense mutations, including G226R, Y296C, R518Q, and S606L [13,33,35], have been observed between Chinese and Japanese (though as yet not identified in Taiwanese) X-linked adrenoleukodystrophy patients, indicating the possibility of inheritance from common ancestors.
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ABCD1 p.Ser606Leu 16996397:126:71
status: NEW124 However, common missense mutations, including G226R, Y296C, R518Q, and S606L [13,33,35], have been observed between Chinese and Japanese (though as yet not identified in Taiwanese) X-linked adrenoleukodystrophy patients, indicating the possibility of inheritance from common ancestors.
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ABCD1 p.Ser606Leu 16996397:124:71
status: NEW
PMID: 21488864
[PubMed]
Kemp S et al: "Mammalian peroxisomal ABC transporters: from endogenous substrates to pathology and clinical significance."
No.
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Comment
259
Two X-ALD causing missense mutations located in the ATP-binding domain of ALDP resulted in either decreased ATP-binding capacity (p.Ser606Leu) or reduced ATPase activity (p.Gly512Ser), when analysed in the context of recombinant nucleotide binding domains (Roerig et al., 2001).
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ABCD1 p.Ser606Leu 21488864:259:132
status: NEW
PMID: 10190819
[PubMed]
Takano H et al: "Mutational analysis and genotype-phenotype correlation of 29 unrelated Japanese patients with X-linked adrenoleukodystrophy."
No.
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Comment
42
Mutations in the ALD Gene That Result in Amino Acid Substitutions or In-frame Deletions* Patient No. Phenotype Mutation† Exon Effect of Mutation‡ Position of Mutation§ Amino Acid Identityሻ Family DataPMP70 mALDRP Pxa1p Amino Acid Deletion G4010 ACALD del.1256-1258 1 del.E291 EAA-like motif E E E CCALD G4011(s) ACALD del.2146-2157¶ 7 del.HILQ587-590 Between Walker A and B# HILE HIVQ YLLK No family history Missense Mutation G4012 CCALD A829G 1 N148S TM3 N N N AMN G1986 CCALD G984A¶ 1 D200N TM4 D D D ACALD G4013 CCALD A1026G¶ 1 N214D TM4 N N N Not available G4014 AMN G1182A 1 G266R Between TM5 and EAA motif G G Non AMN G4015(s) CCALD G1182A 1 G266R Between TM5 and EAA motif G G Non No family history G4016(s) AMN G1197A 1 E271K Between TM5 and EAA motif T E R No family history G4017(s) ACALD A1273G¶ 1 Y296C EAA motif Y Y Y No family history G4018 CCALD A1273G¶ 1 Y296C EAA motif Y Y Y Not available G4019 AMN C1587T¶ 3 R401W Between TM6 and Walker A R R R Asymptomatic carrier G4020 CCALD G1906T¶ 6 G507V Walker A# G G G Not available G4021 CCALD G1939A 6 R518Q Walker A# R R R CCALD G4022 CCALD G1939A 6 R518Q Walker A# R R R Not available G4023 ACALD T2005C¶ 6 F540S Between Walker A and B# F F F Adult asymptomatic carrier G4024(s) CCALD A2017G 6 Q544R Between Walker A and B# Q Q Q No family history G4025 CCALD C2065T 7 S560L Between Walker A and B# P P P Adult asymptomatic carrier G2469(s) ACALD C2157T¶ 7 R591W Between Walker A and B# R R R No family history G2022(s) AMN C2203T 8 S606L Between Walker A and B# S S S No family history G4026 ACALD C2364T 8 R660W C-terminal to Walker B R R R ACALD *ALD indicates adrenoleukodystrophy; ACALD, adult-onset cerebral ALD; CCALD, childhood cerebral ALD; AMN, adrenomyeloneuropathy; (s), apparently sporadic patients; and del., delete.
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ABCD1 p.Ser606Leu 10190819:42:1550
status: NEWX
ABCD1 p.Ser606Leu 10190819:42:1568
status: NEW65 Although each of the remaining 3 mutations (E271K, R401W, and S606L) was identified in only 1 apparently sporadic case of a patient with AMN, a review of the literature indicated that the S606L mutation has been identified in 2 patients with Addison disease only but in no patients with CCALD.33,46 COMMENT MUTATIONS IN THE ALD GENE Because of the low frequency (4%-7%) of large genomic rearrangements in the ALD gene,26,33,34 a detailed nucleotide sequence analysis to detect small nucleotide alterations is required for most cases of ALD.
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ABCD1 p.Ser606Leu 10190819:65:62
status: NEWX
ABCD1 p.Ser606Leu 10190819:65:188
status: NEW85 In the present study, 1 patient with AMN with no family history was identified to have the C2203T (S606L) mutation, although it had previously been reported33,46 to beassociatedexclusivelywiththeAddisondiseaseonlyphe- notype in other studies.
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ABCD1 p.Ser606Leu 10190819:85:99
status: NEW
PMID: 17092750
[PubMed]
Kemp S et al: "X-linked adrenoleukodystrophy: very long-chain fatty acid metabolism, ABC half-transporters and the complicated route to treatment."
No.
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Comment
57
ATP-binding and ATPase activity has been demonstrated for ALDP [17], and Roerig and coworkers demonstrated that two disease causing missense mutations located in the ATP-binding domain resulted in either decreased ATP-binding capacity (Ser606Leu), or reduced ATPase activity (Gly512Ser) [18].
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ABCD1 p.Ser606Leu 17092750:57:236
status: NEW56 ATP-binding and ATPase activity has been demonstrated for ALDP [17], and Roerig and coworkers demonstrated that two disease causing missense mutations located in the ATP-binding domain resulted in either decreased ATP-binding capacity (Ser606Leu), or reduced ATPase activity (Gly512Ser) [18].
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ABCD1 p.Ser606Leu 17092750:56:236
status: NEW
PMID: 17973979
[PubMed]
Kim WS et al: "Role of ATP-binding cassette transporters in brain lipid transport and neurological disease."
No.
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Comment
208
Two ABCD1 mutations associated with X-ALD (Ser606Leu and Gly512Ser) result in decreased ATP binding and reduced ATPase activity, respectively, thereby indicating that an energy dependent transport activity is crucial to the function of ABCD1 (Roerig et al. 2001).
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ABCD1 p.Ser606Leu 17973979:208:43
status: NEW212 Two ABCD1 mutations associated with X-ALD (Ser606Leu and Gly512Ser) result in decreased ATP binding and reduced ATPase activity, respectively, thereby indicating that an energy dependent transport activity is crucial to the function of ABCD1 (Roerig et al. 2001).
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ABCD1 p.Ser606Leu 17973979:212:43
status: NEW
PMID: 21476988
[PubMed]
Zhang X et al: "Conservation of targeting but divergence in function and quality control of peroxisomal ABC transporters: an analysis using cross-kingdom expression."
No.
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153
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.
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ABCD1 p.Ser606Leu 21476988:153:505
status: NEW181 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).
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ABCD1 p.Ser606Leu 21476988:181:148
status: NEW154 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.
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ABCD1 p.Ser606Leu 21476988:154:505
status: NEW182 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).
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ABCD1 p.Ser606Leu 21476988:182:148
status: NEW
PMID: 17542813
[PubMed]
Takahashi N et al: "Adrenoleukodystrophy: subcellular localization and degradation of adrenoleukodystrophy protein (ALDP/ABCD1) with naturally occurring missense mutations."
No.
Sentence
Comment
2
In this study, we chose nine arbitrary mutant human ALDP forms (R104C, G116R, Y174C, S342P, Q544R, S606P, S606L, R617H, and H667D) with naturally occurring missense mutations and examined the intracellular behavior.
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ABCD1 p.Ser606Leu 17542813:2:106
status: NEW3 When expressed in X-ALD fibroblasts lacking ALDP, the expression level of mutant His-ALDPs (S606L, R617H, and H667D) was lower than that of wild type and other mutant ALDPs.
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ABCD1 p.Ser606Leu 17542813:3:92
status: NEW4 Furthermore, mutant ALDP-green fluorescence proteins (S606L and H667D) stably expressed in CHO cells were not detected due to rapid degradation.
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ABCD1 p.Ser606Leu 17542813:4:54
status: NEW9 These results suggest that mutant ALDPs, which have a mutation in the COOH-terminal half of ALDP, including S606L, R617H, and H667D, were degraded by proteasomes after dimerization.
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ABCD1 p.Ser606Leu 17542813:9:108
status: NEW35 We found that mutant ALDPs with the missense mutations in the G116R S342P Q544R R617H H667D Y174C NH2 COOH C sequence Cytosol Membrane Matrix Walker A Walker B S606P, S606L R104C Fig. 1 A putative secondary structure of adrenoleukodystrophy protein.
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ABCD1 p.Ser606Leu 17542813:35:167
status: NEW71 CHO-K1 cells (5 · 105 cells) were cultured in Ham`s F-12 medium with 10% FBS, 70 lg/mL of penicillin, and 140 lg/mL of streptomycin and transfected with 5 lg of pMAM2/ Table 1 Oligonucleotide primer sequences used for the generation of mutant ALDP constructs Construct name Forward primer (5' to 3') (top) R104C GCCTTGGTGAGCTGCACCTTCCTGTCG G116R GCCCGCCTGGACAGAAGGCTGGCC Y174C GCCTACCGCCTCTGCTCCTCCCAG S342P TGGAGCGCCCCGGGCCTGCTCATG Q544R GCATGTTCTACATCCCGCGGAGGCCCTACATGTC S606P AAGGACGTCCTGCCGGGTGGCGAGAAG S606L AAGGACGTCCTGTTGGGTGGCGAGAAG R617H GCAGAGAATCGGCATGGCCCACATGTTCTACCACAGGC H667D TCCCTGTGGAAATACGACACACACTTGCTA The underlined letters indicate the single base mutation leading to an amino acid replacement.
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ABCD1 p.Ser606Leu 17542813:71:513
status: NEW120 However, the ratios of certain His-ALDPs (S606L, R617H, and H667D) were significantly lower than those of wild type His-ALDP and other mutant His-ALDPs.
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ABCD1 p.Ser606Leu 17542813:120:42
status: NEW125 Taken together, it appears His-ALDP (S606L, R617H, and H667D) might be degraded after translation.
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ABCD1 p.Ser606Leu 17542813:125:37
status: NEW127 As shown in Fig. 3d, His-ALDPs (R104C, G116R, S342P, Q544R, S606P, and S606L) exhibited a punctate staining pattern in the cells, which was superimposable on the distribution of catalase in the same cells, suggesting that these mutant His-ALDPs were correctly localized to peroxisomes.
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ABCD1 p.Ser606Leu 17542813:127:71
status: NEW135 In this experiment, we chose R104C, G116R, and S606P (with normal localization in peroxisomes), Y174C (mislocalization), and S606L and H667D (degradation).
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ABCD1 p.Ser606Leu 17542813:135:125
status: NEW141 In contrast, mutant ALDP-GFP (S606L and H667D) was not detected in any subcellular fractions although PMP70 and catalase were recovered in fractions 2 and 3.
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ABCD1 p.Ser606Leu 17542813:141:30
status: NEW150 The fragments were not extractable with 0.1 mol/L sodium carbonate, indicating (a) (c) (b) (d) 400 140 120 100 Expressionratio(%) 80 60 40 His-ALDP R104C G116R Y174C S342P Q544R S606P S606L R617H H667D Catalase 20 0 100 Expressionratio(%) 80 60 40 20 0 350 300 250 200 150 pmol/h/mgprotein 100 50 0 Normal (139T) X-ALD (163T) M ock M ock W ild W ild N one S606L His-ALDP GFP Catalase R 617H H 667D R 104CG 116RY174C S342PQ 544RS606PS606LR 617HH 667D M ock W ildR 104CG 116RY174CS342PQ 544RS606PS606LR 617HH 667D Fig. 3 Expression of wild type and mutant His-adrenoleukodystrophy proteins (ALDPs) in X-linked adrenoleukodystrophy (X-ALD) fibroblasts (163T).
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ABCD1 p.Ser606Leu 17542813:150:184
status: NEWX
ABCD1 p.Ser606Leu 17542813:150:356
status: NEW159 (c) Co-expression of wild or mutant His-ALDP (S606L, R617H, or H667D) with green fluorescence proteins (GFP).
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ABCD1 p.Ser606Leu 17542813:159:46
status: NEW169 Effect of proteasome inhibitors on mutant ALDP The transient and stable expression experiments of mutant ALDPs suggest that mutant ALDPs such as S606L, R617H, H667D, and R104C are degraded by proteases.
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ABCD1 p.Ser606Leu 17542813:169:145
status: NEW180 Degradation of mutant ALDP (S606L) was similarly inhibited by lactacystin, but not by leupeptin, wild R104C G116R ALDP-GFP ALDP PMP70 1 5 10 1 5 10 1 5 10 1 5 10 1 5 10 1 5 10 1 5 10 Non specific ALDP-GFP ALDP PMP70 Non specific ALDP-GFP ALDP PMP70 Non specific Y174C H667D S606P S606L Fig. 4 Subcellular localization of wild type and mutant adrenoleukodystrophy protein (ALDP) -green fluorescence proteins (GFP) in CHO cells.
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ABCD1 p.Ser606Leu 17542813:180:28
status: NEWX
ABCD1 p.Ser606Leu 17542813:180:280
status: NEW181 The mitochondrial and light mitochondrial fraction from CHO cells expressing wild type ALDP and ALDP-GFP or each mutant ALDP-GFP (R104C, G116R, Y174C, S606P, S606L, or H667D) was fractionated by equilibrium density centrifugation on sucrose.
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ABCD1 p.Ser606Leu 17542813:181:158
status: NEW217 S606P and S606L are in the ABC signature motif.
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ABCD1 p.Ser606Leu 17542813:217:10
status: NEW223 Mutant ALDP-GFP (S606L) showed similar results (date not shown).
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ABCD1 p.Ser606Leu 17542813:223:17
status: NEW229 Recently Lie et al. investigated the dimerization of the COOH-terminal half of ALDP by a yeast two-hybrid assay and found that it could dimerize Table 2 Expression and localization of missense ALDPs Mutant ALDP Transient Stable Expressiona Localizationb b-Oxidationc Expressiona Localizationb Wild +++ Px + ++ Px R104Cd , G116R, S342P, Q544R, S606P +++ Px ) ++ Px Y174C +++ mis ) + mis S606L ++ Px ) ) ) R617H, H667D + ) ) ) ) Wild and mutant His-ALDPs or ALDP-GFPs were transiently expressed in X-ALD fibroblasts or stably expressed in CHO cells, respectively.
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ABCD1 p.Ser606Leu 17542813:229:386
status: NEW244 In Fig. 4, the wild type ALDP would dimerize each other or with endogenous ALDP and could be stabilized, but such complex could not be detected on sucrose gradient of CHO cells expressing mutant ALDP (S606L and H667D).
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ABCD1 p.Ser606Leu 17542813:244:201
status: NEW264 Recently, Roerig et al. suggested that mutation of S606L decreases affinity against ATP, although ATP hydrolyzing activity is normal using a NBD fragment expressed in Escherichia coli (Roerig et al. 2001).
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ABCD1 p.Ser606Leu 17542813:264:51
status: NEW265 Our results suggest that the stability of mutant ALDP (S606L) is also affected by (the) mutation.
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ABCD1 p.Ser606Leu 17542813:265:55
status: NEW271 In this study, we found that mutant ALDP (S606L, R617H, and H667D) was degraded by proteasomes together with wild type ALDP.
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ABCD1 p.Ser606Leu 17542813:271:42
status: NEW
PMID: 9242200
[PubMed]
Korenke GC et al: "Variability of endocrinological dysfunction in 55 patients with X-linked adrenoleucodystrophy: clinical, laboratory and genetic findings."
No.
Sentence
Comment
118
Start of Cortisol Start of Age at endocrine Basal increase neurological examination symptoms ACTH cortisol after i.v. ACTH symptoms ALD gene Patient (year-month) (year-month) (pmol/l) (nmol/l) (nmol/l) (year-month) mutation 1a 9-05 6-08 223 179 0 7-02 cALD 2252-10 G-A 1b 13-03 9-05 41 331 41 9-05 cALD 2252-10 G-A 2a 26-01 - 6 386 221 - R418W 2b 27-10 - 6 461 229 11-00 cALD R418W 3a 11-06 6-00 536 138 8 - S606L 3b 11-06 6-00 423 143 11 10-10 cALD S606L 4a 4-04 2-09 >330 127 141 - R617C 4b 5-06 3-11 >330 97 50 - R617C 4c 7-10 5-05 >330 50 6 6-03 cALD R617C 5a 10-06 - 8 433 290 8-02 cALD del 1257-9 5b 17-07 - 9 637 629 - del 1257-9 6a 27-00 24-00 >320 30 11 18-03 AMN L107P 6b 32-10 - 47 279 185 19-01 AMN L107P 7a 34-08 - 48 469 350 24-00 AMN R418W 7b 36-05 - 43 486 201 28-00 AMN R418W 8a 14-05 - 33 287 94 - 1801 del AG 8b 16-10 - 14 348 149 - 1801 del AG 9a 13-07 5-10 1094 36 6 - 740 del G 9b 157-04 1241 63 11 - 740 del G cALD, cerebral adrenoleucodystrophy; AMN, adrenomyeloneuropathy; -, no symptoms.
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ABCD1 p.Ser606Leu 9242200:118:408
status: NEWX
ABCD1 p.Ser606Leu 9242200:118:450
status: NEW
PMID: 12530690
[PubMed]
Gartner J et al: "Functional characterization of the adrenoleukodystrophy protein (ALDP) and disease pathogenesis."
No.
Sentence
Comment
41
The mutant constructs included missense mutations of patients with X-ALD in the nucleotide binding fold regions Walker A and 19mer (ALDP-NBF-G512S, ALDP-NBF-Q544R, ALDP-NBF-P560L, ALDP-NBF-R591Q, ALDP-NBF-S606L, and ALDP-NBF-D629H) and corresponding mutations in another ABC transporter in the peroxisome membrane, the 70 kDa peroxisomal membrane protein (PMP70; PMP70-NBF-G478R, PMP70- NBF-S572I).
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ABCD1 p.Ser606Leu 12530690:41:205
status: NEW60 In contrast, the patient mutation S606L alters ATP binding affinity but has no effect on the overall ATPase activity.
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ABCD1 p.Ser606Leu 12530690:60:34
status: NEW81 In contrast, missense mutations included in the second group like S606L in the ALD gene reduce ATP-binding but do not have any effect on the overall ATPase activity.
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ABCD1 p.Ser606Leu 12530690:81:66
status: NEW
PMID: 7668254
[PubMed]
Watkins PA et al: "Altered expression of ALDP in X-linked adrenoleukodystrophy."
No.
Sentence
Comment
176
In 11 patients, missense mutations that occurred throughout the protein were found: within the transmembrane domains (patients 1, 3, and 4), within the ATP-binding domain (patients 8-12), and on either side of the ATP-binding Table 3 Mutational Analysis of the ALD Gene in IS Unrelated Patients ALDP Patient Phenotype Mutation Consequence Immunoreactivity 1 .................. CALD 825 A-GG K276E + 2.................. AMN 870-2AGAGE291,& 3 .................. CALD 872 G-C E291D 4 .................. AMN 1023 T-IC S342P+ 5 .................. AMN 1166 G-C R389H + 6 .................. CALD 1201 G-AA R401Q + 7 ........ CALD 1415-6 AAG FS@472 8 ........ AMN 1771 G-AA R591Q + 9 ........ Addison 1817 C-T S606L + 10 ................ AMN 1850 G-AA R617H 11 ................ CALD 1876 G-AA A626T 12 ................ Fetus 1884 G-C D629H + 13 ................ CALD 1932 C-UT Q645X 14 ................ AMN 1978 C-OT R660W 15 ........ AMN AExon7-10 Null Mutations in the ALD gene were determined, as described in Methods, in 15 of the ALD patients reported in table 2.
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ABCD1 p.Ser606Leu 7668254:176:705
status: NEW178 In 11 patients, missense mutations that occurred throughout the protein were found: within the transmembrane domains (patients 1, 3, and 4), within the ATP-binding domain (patients 8-12), and on either side of the ATP-binding Table 3 Mutational Analysis of the ALD Gene in IS Unrelated Patients ALDP Patient Phenotype Mutation Consequence Immunoreactivity 1 .................. CALD 825 A-GG K276E + 2 .................. AMN 870-2 AGAG E291,& 3 .................. CALD 872 G-C E291D 4 .................. AMN 1023 T-IC S342P + 5 .................. AMN 1166 G-C R389H + 6 .................. CALD 1201 G-AA R401Q + 7 ........ CALD 1415-6 AAG FS@472 8 ........ AMN 1771 G-AA R591Q + 9 ........ Addison 1817 C-T S606L + 10 ................ AMN 1850 G-AA R617H 11 ................ CALD 1876 G-AA A626T 12 ................ Fetus 1884 G-C D629H + 13 ................ CALD 1932 C-UT Q645X 14 ................ AMN 1978 C-OT R660W 15 ........ AMN AExon7-10 Null Mutations in the ALD gene were determined, as described in Methods, in 15 of the ALD patients reported in table 2.
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ABCD1 p.Ser606Leu 7668254:178:709
status: NEW
PMID: 8773611
[PubMed]
Korenke GC et al: "Cerebral adrenoleukodystrophy (ALD) in only one of monozygotic twins with an identical ALD genotype."
No.
Sentence
Comment
85
The S606L mutation reported here has already been described in a patient with ALD and Addison's disease [l11.
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ABCD1 p.Ser606Leu 8773611:85:4
status: NEW
No.
Sentence
Comment
47
ALDP is an integral peroxisomal membrane protein with the nucleotide-binding domain located toward the cytoplasmic surface of the peroxisomal membrane (14).ATP-binding andATPase activity has been demonstrated for ALDP (71), and Roerig et al demonstrated that two disease causing missense mutations located in the ATP-binding domain resulted in either decreased ATP-binding capacity (p.Ser606Leu) or reduced ATPase activity (p.Gly512Ser) (64).
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ABCD1 p.Ser606Leu 20626744:47:385
status: NEW
PMID: 7581394
[PubMed]
Kok F et al: "Mutational analysis of patients with X-linked adrenoleukodystrophy."
No.
Sentence
Comment
131
3' deletion 3' deletion 3' deletion 3' deletion R104C A141T R152C R182P Frameshift at AA 231 G277W R389H Spl mutation at AA 408 Q466 stop Frameshift at AA 470 Frameshift at AA 470 Frameshift at AA 472 Frameshift at AA 472 Frameshift at AA 472 Frameshift at AA 472 Frameshift at AA 472 Frameshift at AA 472 Frameshift at AA 472 G512S M566K S606L L516L R617H R660W - - Exons 3-10 Exons 7-10 Exons 8-10 Exons 7-10 33 Anglos 5 Scott 8 Anglos 7 Anglos 11 Jewish 36 Irish 51 Italian 37 Filipino 28 Anglos 23 Anglos 11 Anglos 8 Anglos 40 Italian 22 German 4 Anglos 5 black 8 Anglos 31 Anglos 10 Anglos 28 Anglos 22 Italian 8 German 35 German 7 Hispanic 28 German 24 Anglos 18 Jewish 9 Hispanic AMNa C E R ~ Cer Add' Cer AMN AMN AMN AMN Cer Cer Cer Add AMN AMN Cer Cer Cer AMN Add AMN AMN Cer AMN Cer AMN AMN AMN 5 Cer,AMN,Add 4 Cer,AMN 1 Cer 5 Cer,AMN,Add 1 4 2 1 2 2 5 Adopted 5 2 15 1 13 2 2 1 Cer AMN AMN,Add AMN Cer,AMN Cer,AMN Cer,AMN,Add ?
X
ABCD1 p.Ser606Leu 7581394:131:339
status: NEW234 Four mutations were found within the ATP bind- ing domain, one in Walker A (G512S), one in Walker B (R617H), one in the highly conserved sequence preceding Walker B (S606L),and one in the middle of the domain.
X
ABCD1 p.Ser606Leu 7581394:234:169
status: NEW
PMID: 11248239
[PubMed]
Roerig P et al: "Characterization and functional analysis of the nucleotide binding fold in human peroxisomal ATP binding cassette transporters."
No.
Sentence
Comment
5
Mutations in conserved residues of the nucleotidases (PMP70: G478R, S572I; ALDP: G512S, S606L) altered ATPase activity.
X
ABCD1 p.Ser606Leu 11248239:5:88
status: NEW24 For the mutant constructs we selected X-ALD patient mutations in highly conserved residues in the Walker A and 19-mer region of the NBF of ALDP (G512S and S606L) and the corresponding PMP70 mutations (G478R and S572I).
X
ABCD1 p.Ser606Leu 11248239:24:155
status: NEW85 Additionally, we changed the conserved serine in the 19-mer motif of PMP70 and ALDP to isoleucine (S572I) and leucine (S606L), respectively.
X
ABCD1 p.Ser606Leu 11248239:85:119
status: NEW88 The K-subunit of L-galactosidase (L-Protein Synthetic oligonucleotide Mutation ALDP 5P-CCCCAATGGCTGCAGCAAGAGCTCCC-3P G512S 5P-GGATCCGGACAGGGAGCTCTTGCTGCAGC-3P ALDP 5P-ACTGGAAGGACGTCCTGTTGGG-3P S606L 5P-CGCCACCCAACAGGACGTCCTTCC-3P PMP70 5P-GGCTGCAGAAAGAGTTCACTTTTCCG-3P G478R 5P-GGCCATAATTCACCAAGAACACGGAAA AGTGAACTCTTTCTG-3P PMP70 5P-GACGTACTCATTGGTGGAG-3P S572I 5P-CCACCAATGAGTACGTCCATCCAATCC-3P gal) in fusion with the MBP was used as a control.
X
ABCD1 p.Ser606Leu 11248239:88:193
status: NEW117 G512S and S606L cause X-ALD.
X
ABCD1 p.Ser606Leu 11248239:117:10
status: NEW136 The S606L and G478R mutants have a decreased ATP binding a¤nity while the G512S and S572I mutants decrease the maximum velocity of ATPase activity.
X
ABCD1 p.Ser606Leu 11248239:136:4
status: NEW171 In contrast, the X-ALD patient mutation S606L alters ATP binding a¤nity but has no e¡ect on the overall ATPase activity.
X
ABCD1 p.Ser606Leu 11248239:171:40
status: NEW172 Because the carboxyl-terminal halves of ALDP, PMP70 and ALDR are involved in protein dimerization and interactions with other protein motifs like the transmembrane domain [28], it is possible that the S606L missense mutation in ALDP exerts its disease causing e¡ect in this manner.
X
ABCD1 p.Ser606Leu 11248239:172:40
status: NEWX
ABCD1 p.Ser606Leu 11248239:172:201
status: NEW182 Previous studies on MDR suggest that the ATPase activity of the native protein Table 1 Kinetic parameters of ATPase activity in wild type and mutant ALDP and PMP70 NBF fusion proteins Fusion protein KM (WM) Vmax (nmol/Wmol NBF/min) Speci'c activity (1033 U/mg) ALDP (wild type) 11.5 þ 0.97 641.9 þ 10.7 10.0 þ 0.17 ALDP (G512S) 17.9 þ 1.23 279.3 þ 4.2 4.4 þ 0.06 ALDP (S606L) 45.6 þ 2.40 666.0 þ 8.7 10.4 þ 0.14 PMP70 (wild type) 8.2 þ 0.52 580.8 þ 6.7 9.0 þ 0.10 PMP70 (G478R) 161.8 þ 34.40 641.2 þ 28.2 10.0 þ 0.44 PMP70 (S572I) 9.9 þ 0.82 298.1 þ 4.7 4.6 þ 0.07 The kinetic data of all fusion proteins are mean values and standard deviations of 15^20 measurements at various protein concentrations using at least two distinct protein preparations.
X
ABCD1 p.Ser606Leu 11248239:182:399
status: NEW86 Additionally, we changed the conserved serine in the 19-mer motif of PMP70 and ALDP to isoleucine (S572I) and leucine (S606L), respectively.
X
ABCD1 p.Ser606Leu 11248239:86:119
status: NEW89 The K-subunit of L-galactosidase (LProtein Synthetic oligonucleotide Mutation ALDP 5P-CCCCAATGGCTGCAGCAAGAGCTCCC-3P G512S 5P-GGATCCGGACAGGGAGCTCTTGCTGCAGC-3P ALDP 5P-ACTGGAAGGACGTCCTGTTGGG-3P S606L 5P-CGCCACCCAACAGGACGTCCTTCC-3P PMP70 5P-GGCTGCAGAAAGAGTTCACTTTTCCG-3P G478R 5P-GGCCATAATTCACCAAGAACACGGAAA AGTGAACTCTTTCTG-3P PMP70 5P-GACGTACTCATTGGTGGAG-3P S572I 5P-CCACCAATGAGTACGTCCATCCAATCC-3P gal) in fusion with the MBP was used as a control.
X
ABCD1 p.Ser606Leu 11248239:89:192
status: NEW118 G512S and S606L cause X-ALD.
X
ABCD1 p.Ser606Leu 11248239:118:10
status: NEW137 The S606L and G478R mutants have a decreased ATP binding a&#a4;nity while the G512S and S572I mutants decrease the maximum velocity of ATPase activity.
X
ABCD1 p.Ser606Leu 11248239:137:4
status: NEW173 Because the carboxyl-terminal halves of ALDP, PMP70 and ALDR are involved in protein dimerization and interactions with other protein motifs like the transmembrane domain [28], it is possible that the S606L missense mutation in ALDP exerts its disease causing e&#a1;ect in this manner.
X
ABCD1 p.Ser606Leu 11248239:173:201
status: NEW183 Previous studies on MDR suggest that the ATPase activity of the native protein Table 1 Kinetic parameters of ATPase activity in wild type and mutant ALDP and PMP70 NBF fusion proteins Fusion protein KM (WM) Vmax (nmol/Wmol NBF/min) Speci'c activity (1033 U/mg) ALDP (wild type) 11.5 &#fe; 0.97 641.9 &#fe; 10.7 10.0 &#fe; 0.17 ALDP (G512S) 17.9 &#fe; 1.23 279.3 &#fe; 4.2 4.4 &#fe; 0.06 ALDP (S606L) 45.6 &#fe; 2.40 666.0 &#fe; 8.7 10.4 &#fe; 0.14 PMP70 (wild type) 8.2 &#fe; 0.52 580.8 &#fe; 6.7 9.0 &#fe; 0.10 PMP70 (G478R) 161.8 &#fe; 34.40 641.2 &#fe; 28.2 10.0 &#fe; 0.44 PMP70 (S572I) 9.9 &#fe; 0.82 298.1 &#fe; 4.7 4.6 &#fe; 0.07 The kinetic data of all fusion proteins are mean values and standard deviations of 15^20 measurements at various protein concentrations using at least two distinct protein preparations.
X
ABCD1 p.Ser606Leu 11248239:183:393
status: NEW
PMID: 8040304
[PubMed]
Fanen P et al: "Identification of mutations in the putative ATP-binding domain of the adrenoleukodystrophy gene."
No.
Sentence
Comment
48
Asterisks indicate the posi- *l I U13 | tion of the four missense mutations in ALD protein (ALDP): R518W substitution occurs at the same amino acid position as in the CFTR mutant S1255P, and S606L at the same position as in the S5491 or S549R CFTR mutants; R617C and R617H have no equivalents in CFTR mutants.
X
ABCD1 p.Ser606Leu 8040304:48:191
status: NEW88 The C2203 -+ T (Ser6O6 -+ Leu or S606L) mutation was identified in a patient who had Addison's disease without neurologic involvement at 20 years.
X
ABCD1 p.Ser606Leu 8040304:88:33
status: NEW127 Three of them (S606L, R617C, and R617H) involve invariant residues in all ABC proteins studied so far (Fig. 1).
X
ABCD1 p.Ser606Leu 8040304:127:15
status: NEW143 The CFTR mutants S5491 and S549R, that involve the same amino acid position as in the ALD mutant S606L, fail to produce mature CFTR and therefore prevent trafficking to the correct cellular localization ( 12).
X
ABCD1 p.Ser606Leu 8040304:143:97
status: NEW145 Mutations Detected in the ALD Gene Name Nucleotide change Effect on coding sequence Exon Clinical phenotype R464X C - T at 1776 Arg - Stop at 464 4 AMN* 1937delC Deletion of C at 1937 Frameshift 6 cerebral ALD R518W CT at 1938 Arg-Trpat 518 6 AMN 2020 + I G - A G - A at 2020 + 1 5' splice signal Intron 6 ACMN2 2177delTA Deletion of TA at 2177 Frameshift 8 cerebral ALD S606L C - T at 2203 Ser - Leu at 606 8 Addison 2204delG Deletion of G at 2204 Frameshift 8 Addison R617C C - T at 2235 Arg - Cys at 617 8 cerebral ALD R617H G - A at 2236 Arg - His at 617 8 ACMN * Adrenomyeloneuropathy; tadrenomyeloneuropathy with cerebral involvement.
X
ABCD1 p.Ser606Leu 8040304:145:371
status: NEW
PMID: 16087056
[PubMed]
Pan H et al: "ABCD1 gene mutations in Chinese patients with X-linked adrenoleukodystrophy."
No.
Sentence
Comment
94
RFLP Associated Phenotype Missense mutations A30 1 421 GϾA A141T CCALD A23 1 545 GϾC R182P CCALD A14 1 796 GϾA G266R‡ CCALD A18 1 847 CϾG H283D†§ - Nsp I CCALD A32 1 871 GϾA E291K‡ CCALD A28 1 887 AϾG Y296C ACALD A21 2 1028 GϾT G343V*‡§ - Ava I ACALD A20 2, 3 1047 CϾA V349V*§ ϩ Rsa I CCALD 1210 TϾC S404P†‡ A6 6 1526 AϾT N509I†‡ CCALD A26 6 1529 GϾA G510D*§ - Bg1 I ACALD A1 6 1552 CϾG R518G†‡§ - Msp I CCALD A24 6 1548 GϾA L516L‡ CCALD 1553 GϾA R518Q‡ A10 6 1553 GϾA R518Q‡ CCALD A7 6 1559 TϾA L520Q CCALD A12 7 1661 GϾA R554H‡ CCALD A19 7 1667 AϾG Q556R‡ AMN A16 8 1814 TϾA L605Q*§ ϩ BstX I CCALD A17 8 1817 CϾT S606L CCALD A2 8 1849 CϾT R617C‡ AO A15 8 1849 CϾG R617G CCALD Nonsense mutations A11 1 396 GϾA W132X‡ CCALD A3 1 726 GϾA W242X CCALD A34 4 1390 CϾT R464X‡ CCALD A8 8 1785 GϾA W595X‡ CCALD Frameshift mutations A29 1 385 ins G fs R128* ACALD A27 2 937 del C fs D312 CCALD A13 5 1415 del AG fs E471 ACALD A22 6 1603 del CC fs P534* CCALD Amino acid insertion A33 1 240-241ins9 R80-L81insPAA* CCALD Splicing defect A5 IVS1 IVS1 ϩ1 gϾt CCALD A31 IVS3 IVS3 ϩ2 cϾt CCALD A25 IVS5 IVS5 -6 delc†‡ ACALD Synonymous mutation A4 2, 6 1047 CϾA V349V‡ CCALD 1548 GϾA L516L‡ A9 2, 6 1047 CϾA V349V‡ CCALD 1548 GϾA L516L‡ * The mutation was novel.
X
ABCD1 p.Ser606Leu 16087056:94:882
status: NEW95 RFLP Associated Phenotype Missense mutations A30 1 421 Gb0e;A A141T CCALD A23 1 545 Gb0e;C R182P CCALD A14 1 796 Gb0e;A G266Rߥ CCALD A18 1 847 Cb0e;G H283Dߤ&#a7; afa; Nsp I CCALD A32 1 871 Gb0e;A E291Kߥ CCALD A28 1 887 Ab0e;G Y296C ACALD A21 2 1028 Gb0e;T G343V*ߥ&#a7; afa; Ava I ACALD A20 2, 3 1047 Cb0e;A V349V*&#a7; af9; Rsa I CCALD 1210 Tb0e;C S404Pߤߥ A6 6 1526 Ab0e;T N509Iߤߥ CCALD A26 6 1529 Gb0e;A G510D*&#a7; afa; Bg1 I ACALD A1 6 1552 Cb0e;G R518Gߤߥ&#a7; afa; Msp I CCALD A24 6 1548 Gb0e;A L516Lߥ CCALD 1553 Gb0e;A R518Qߥ A10 6 1553 Gb0e;A R518Qߥ CCALD A7 6 1559 Tb0e;A L520Q CCALD A12 7 1661 Gb0e;A R554Hߥ CCALD A19 7 1667 Ab0e;G Q556Rߥ AMN A16 8 1814 Tb0e;A L605Q*&#a7; af9; BstX I CCALD A17 8 1817 Cb0e;T S606L CCALD A2 8 1849 Cb0e;T R617Cߥ AO A15 8 1849 Cb0e;G R617G CCALD Nonsense mutations A11 1 396 Gb0e;A W132Xߥ CCALD A3 1 726 Gb0e;A W242X CCALD A34 4 1390 Cb0e;T R464Xߥ CCALD A8 8 1785 Gb0e;A W595Xߥ CCALD Frameshift mutations A29 1 385 ins G fs R128* ACALD A27 2 937 del C fs D312 CCALD A13 5 1415 del AG fs E471 ACALD A22 6 1603 del CC fs P534* CCALD Amino acid insertion A33 1 240-241ins9 R80-L81insPAA* CCALD Splicing defect A5 IVS1 IVS1 af9;1 gb0e;t CCALD A31 IVS3 IVS3 af9;2 cb0e;t CCALD A25 IVS5 IVS5 afa;6 delcߤߥ ACALD Synonymous mutation A4 2, 6 1047 Cb0e;A V349Vߥ CCALD 1548 Gb0e;A L516Lߥ A9 2, 6 1047 Cb0e;A V349Vߥ CCALD 1548 Gb0e;A L516Lߥ * The mutation was novel.
X
ABCD1 p.Ser606Leu 16087056:95:885
status: NEW
PMID: 17202797
[PubMed]
Takahashi N et al: "[Adrenoleukodystrophy: structure and function of ALDP, and intracellular behavior of mutant ALDP with naturally occurring missense mutations]."
No.
Sentence
Comment
8
We found that mutant ALDP (S606L, R617H, and H667D) was degraded together with wild-type ALDP by proteasomes.
X
ABCD1 p.Ser606Leu 17202797:8:27
status: NEW19 S606P and S606L are in ABC signature motif.
X
ABCD1 p.Ser606Leu 17202797:19:10
status: NEW28 ミスセンス変異を持つ ALDP の細胞内動態 ―一過性発現による解析 ALD 患者の持つ変異 ALDP の機能,細胞内局在 性,細胞内における安定性を解析することは, ALDP の各ドメインの機能を知る上で有用な情報 を提供すると思われる．特にミスセンス変異は,た った 1 つのアミノ酸変異による異常であるので特に 興味深い．われわれは ALD 患者で報告されている ミスセンス変異の中から,TMD から 4 つ(R104C, G116R, Y174C, S342P),NBD から 4 つ(Q544R, S606P, S606L, R617H),C 末端部位から 1 つ (H667D)を任意に選び(Fig. 1),その機能と細胞 内動態を解析した．これらの実験は,大学院シンポ ジウムで報告したので,詳しく述べたいと思う． ALDP はペルオキシソームにおける極長鎖脂肪 酸の b 酸化に関与していることが知られている． 実際に ALD 患者由来の繊維芽細胞では極長鎖脂肪 酸の b 酸化活性が正常な線維芽細胞と比べて約 50 ―70%程度減少している．そこで野生型及び変異型 ALDP の機能を確認するため,ALDP を発現して いない ALD 患者由来線維芽細胞に,N 末端に His タグを付加した野生型と変異型 ALDP を一過性に 発現し,［1-14 C]lignoceric acid を基質として極長鎖 脂肪酸 b 酸化活性の測定を行った．その結果, ALDP 欠損線維芽細胞の極長鎖脂肪酸 b 酸化活性 は,正常細胞の約 50%まで減少していたが,野生 型 His-ALDP を発現させると正常と同程度にまで 活性が回復した．このことから発現させた野生型 His-ALDP は ALDP と同等の機能を持つことが確 認された．一方,9 種類のミスセンス変異 ALDP を発現した線維芽細胞では極長鎖脂肪酸 b 酸化活 性の増加は認められなかった．よって,これらのミ スセンス変異 ALDP は機能を欠くことが確認され た． ついで,野生型及び変異型 His-ALDP を発現し た ALD 患者線維芽細胞を回収し,変異型 ALDP の発現量を immunoblotting により定量化し解析し た(Table 1)．なお ALDP の発現量は,ペルオキ シソームの指標酵素であるカタラーゼの発現量で補 正した．その結果,変異型 ALDP(R104C, G116R, Y174C, S342P, Q544R, S606P)は,野生型とほぼ 同程度の発現量を示した．一方,変異型 ALDP (S606L, R617H, H667D)では発現量が野生型の発 167 Table 1.
X
ABCD1 p.Ser606Leu 17202797:28:229
status: NEWX
ABCD1 p.Ser606Leu 17202797:28:1388
status: NEWX
ABCD1 p.Ser606Leu 17202797:28:5950
status: NEW29 Expression and Localization of Missense ALDPs Mutant Transient Stable Expression Localization b-Oxidation Expression Localization Wild Z Px ＋ Z Px R104C, G116R S342P, Q544R S606P Z Px － Z Px Y174C Z mis － Z mis S606L ＋ Px － － － R617H ± － － na na H667D ＋ － － － － Wild and Mutant His-ALDPs or ALDP-GFPs were transiently expressed in X-ALD ˆbroblasts and stably expressed in CHO cells, respectively.
X
ABCD1 p.Ser606Leu 17202797:29:232
status: NEW36 1 現量と比べて約 50%程度減少していた．なお,各 ALDP ポジティブの細胞は約 30%程度であり,各 細胞間での発現効率に有意な差は認められなかっ た．このことから,ALDP の発現量が減少してい た 3 つの変異 ALDP は細胞内での安定性が低下し ていると推察された．また興味深いことに S606P と S606L は同じ部位の変異にも係わらず,置換し たアミノ酸によって発現量には差が認められた． ついで,変異型 His-ALDP の細胞内局在を蛍光 抗体法で確認した．変異型 ALDP(R104C, G116R, S342P, Q544R, S606P, S606L)では ALDP がカタ ラーゼの局在と一致したことから,正常にペルオキ シソームへ局在していることが確認された．一方, 変異型 ALDP(Y174C, H667D)では局在が一致せ ず,ALDP が他の細胞内小器官へ間違って輸送さ れていると考えられた．変異型 ALDP(R617H) では ALDP の発現が認められなかった．変異型 ALDP(R104C, G116R, S342P, Q544R, S606P)で は野生型とほぼ同程度のタンパク量が発現し,ペル オキシソームへの局在も確認されたので,これらの 変異型 ALDP は合成されたのちに正常にペルオキ シソームに運ばれるが,ペルオキシソーム膜におい てその機能(ATP 結合・加水分解若しくは基質輸 送)に異常を持つことが推察された．特に R104C, G116R, S342P は TMD に存在することから ALDP の基質輸送能が変化していると考えられる．一方, NBD に存在する Q544R, S606P は ATP 結合・加水 分解に影響を与えている可能性が考えられる．また S606P, S606L は変異が同じ部位でも構造的に安定 性が異なっていた．Roerig らは S606L の変異型 ALDP は,ATP との親和性が低下している一方で ATP 加水分解は正常に行われていると報告してい る．29) このことは ALDP と ATP の親和性が ALDP の安定性にも影響を及ぼしている可能性を示してい る．S606L と S606P の安定性の違いと機能の関係 は ALDP の機能を知る上でも興味深い点であり, 今後さらに検討を行う必要がある．一方,Y174C の変異型 ALDP は正常に発現するにも係わらず, ペルオキシソームへ局在せず他の細胞内小器官へミ スターゲッティングした．これまでにペルオキシ ソームへの局在化シグナルを欠くペルオキシソーム 膜タンパク質は,非特異的にミトコンドリアや小胞 体に移行することが知られている．30,31) よって, ALDP の TMD2―3 の間のループは,ペルオキシ ソームへの局在化に重要な役割を果たしている可能 性が推察される．Pex19p 存在化での in vitro タン パク質翻訳系において,ALDP(Y174C)は Pex19p に結合できるので,ALDP の N 末端 67―164 に存在するペルオキシソーム移行に係わる領域が ALDP の何らかの構造変化によってマスクされる のかもしれない． 5.
X
ABCD1 p.Ser606Leu 17202797:36:949
status: NEWX
ABCD1 p.Ser606Leu 17202797:36:1539
status: NEWX
ABCD1 p.Ser606Leu 17202797:36:4121
status: NEW43 168 Vol. 127 (2007) ALDP については ALDP の C 末端に GFP(green ‰uorescent protein)を融合させた変異 ALDP-GFP を発現させるとともに,野生型ヒト ALDP を CHO 細胞に共 発現させた(CHO 細胞にも 内在性の ALDP が発現しているが,本実験に用いた抗体が 交差しないため,ヒト ALDP を発現させた)．GFP 融合タンパク質は内在性のタンパク質との区別が容 易であること,安定発現細胞の取得が容易にできる ことなどの利点がある．この実験では,ペルオキシ ソームに正常に輸送される変異型 ALDP(R104C, G116R, S606P),ペルオキシソームに局在しない変 異型 ALDP(Y174C),発現量が低下している変異 型 ALDP(H667D)を選んだ．これら安定過剰発 現細胞における ALDP-GFP の細胞内局在性をみる と,その分布は一過性発現させた His-ALDP と同 様であった． 得られた安定発現細胞よりオルガネラ粗分画を調 製し,ショ糖密度勾配遠心分離法により各フラクシ ョンに分けたのち,SDS-PAGE 及び immunoblotting により変異 ALDP の局在について解析を行っ た(Fig. 3)．ペルオキシソームマーカーとしてペ ルオキシソーム膜タンパク質である PMP70 とペル オキシソームの主要なマトリックスタンパク質であ るカタラーゼを用いた．野生型 ALDP を安定過剰 発現している細胞において,カタラーゼ活性並びに PMP70 が主としてフラクション 3 及び 4 に存在す ることより,この分画にペルオキシソームが回収さ れたことが示唆された．また約 110 kDa の分子サ イズを持つ ALDP-GFP 並びに 83 kDa の野生型 ALDP は,ペルオキシソームマーカーとほぼ同じ 分布を示していたことからペルオキシソームに局在 していることが示唆された．また変異型 ALDP-GFP(G116R, S606P)も同様の分布を示した． 一方,変異型 ALDP-GFP(H667D)を安定過剰 発現している細胞の場合は,ALDP-GFP のバンド は検出されなかった(Fig. 3)．興味深いことに, PMP70 は 検 出 さ れ た が , 共 発 現 さ せ た 野 生 型 ALDP のバンドも検出されなかった．また変異型 ALDP(S606L)についても同様であった．これら の 結 果 は , 変 異 型 ALDP ( H667D, S606L ) は PMP70 とではなく,野生型 ALDP と複合体を形成 し,両者が分解される可能性を示唆している． ABC タンパク質の機能発現に重要である TMD や NBD 以外の C 末端部位での変異がタンパク質の安 定性に影響を及ぼすことは興味深い．ALDP の C 末端部位である 600―700 アミノ酸での変異が X-ALD を引き起こす頻度が高いことから,ALDP の C 末端部位はタンパク質の安定性に重要な役割を担 っている可能性がある．Liu らは ALDP のダイ マー化には C 末端部位(AA.631―745)が重要で あると報告している．13) H667D や S606L のような 変異は,それ自身あるいは野生型 ALDP とミスフ ォールドしたダイマーを形成し,異常タンパク質と して認識され分解されると考えられる．一方,変異 型 ALDP-GFP(R104C)はペルオキシソーム分画 に回収されるものの,フラグメント化していること 169 Fig. 4.
X
ABCD1 p.Ser606Leu 17202797:43:5513
status: NEWX
ABCD1 p.Ser606Leu 17202797:43:5719
status: NEWX
ABCD1 p.Ser606Leu 17202797:43:7320
status: NEW49 変異型 ALDP の分解過程の解析 新生タンパク質が正し;044;フォールディングを受け ることは,そのタンパク質の正常な機能発現のため に必須である．遺伝子変異などが存在すると,タン パク質がミスフォールディングされる．このミスフ ォールドタンパクが細胞外へ分泌されたり,細胞内 に蓄積したりすると生体にとって極めて有害になる ため,このようなタンパクはプロテアソーム,リソ ソーム等によって迅速に分解される．ちなみに,嚢 胞性線維症の原因タンパク質 CFTR は細胞膜イオ ンチャネルとして機能する ABC タンパク質である が,変異 CFTR は小胞体膜からプロテアソームに リクルートされ分解されることが報告されてい る．32,33) しかしながら,変異型 ALDP を始めとし て,ペルオキシソーム膜タンパク質についての解析 はほとんど行われていない． 変異型 ALDP の一過性発現と安定過剰発現実験 より,ALDP(S606L, R617H, H667D, R104C)は, プロテアーゼにより分解されていると推定された． そこで,ALDP-GFP(H667D)を発現している CHO 細胞に各種プロテアーゼ阻害剤を処理し,解 析を行った．その結果,プロテアソーム阻害剤であ る lactacystin を処理した細胞では ALDP-GFP 及び ALDP の バ ン ド が 出 現 し た ( Fig. 4 )． 一 方 , leupeptin, AEBSF, E64d には効果がなかった．ま た他のプロテアソーム阻害剤である MG132 も有効 であった．さらにプロテアソーム阻害剤により分解 を逃れた変異型 ALDP-GFP(H667D)の細胞内局 在を蛍光抗体法で観察すると,ペルオキシソームに 局在していることが確認された．一方,変異型 ALDP(R104C)のフラグメント化は上記プロテアー ゼ処理では阻害されなかった． さらに ALD 患者由来細胞の内因性変異 ALDP の分解とプロテアソーム分解系の関与について確認 するため,変異型 ALDP(R617H)を持つ患者由 来線維芽細胞を用いてタンパク分解の阻害実験を行 った．その結果,lactacystin と MG132 処理により, ALDP のバンドが出現した．以上の結果より,ペ ルオキシソーム膜上にはミスフォールドしたタンパ ク質を認識する仕組みが存在し,プロテアソーム及 び他のプロテアーゼを介して排除していることが示 唆された． 一方,山田らは ALD 患者線維芽細胞を［35 S］メチ オニンでパルスチェイスすることにより,変異型 ALDP(G512S, R660W)の分解が E-64 と leupepu- tin により抑制されることを報告している．34) 彼ら の実験ではプロテアソーム阻害剤については実験し ていないので,プロテアソームの関与は不明である が,変異型 ALDP の分解には,複数のプロテアー ゼが関与している可能性がある． 7.
X
ABCD1 p.Ser606Leu 17202797:49:174
status: NEWX
ABCD1 p.Ser606Leu 17202797:49:2770
status: NEW52 Some mutant ALDPs (R104C, G116R, S342P, Q544R and S606P) are normally inserted into the peroxisomal membrane, and others were mislocalized (Y174C) or degraded by proteasome (S606L, R617H and H667D).
X
ABCD1 p.Ser606Leu 17202797:52:174
status: NEW53 170 Vol. 127 (2007) その結果より,ミスセンス変異 ALDP は以下に 示すように 4 種類の細胞内動態を持つことが示され た(Fig. 5)．1) 野生型と同様にペルオキシソーム に 局 在 す る が そ の 機 能 が 阻 害 さ れ て い る 変 異 (R104C, G116R, S342P, Q544R, S606P),2) ペル オキシソームへの局在化に障害がある変異(Y174C), 3) 変異によりタンパク質の安定性が低下しプロテ アソームでの分解を受けるが,一部ペルオキシソー ムに局在する変異(S606L),4) 変異によりタンパ ク質の安定性が低下しプロテアソームで選択的に分 解を受け,細胞内でほとんど確認できない変異 (R617H, H667D)の 4 種類のパターンである． 発現量も局在化も正常な変異では,ABC タンパ ク質としての機能に直接関与している機能ドメイン の障害が起こっていると推察される．この中で G116R, S342P は TMD に位置しており,基質の認 識や輸送に障害があると推察される．また Q544R, S606P は ATP と の 結 合 ・ 加 水 分 解 に 関 与 す る NBD に位置している．このような変異は,ALDP の ABC タンパク質としての機能を解析するために 有益と考えられる． 発現量は正常だが局在化に異常が認められた Y174C は,TMD2 と 3 の間のループ 2 に位置して おり,この領域が ALDP のペルオキシソームへの ターゲッティングに必要であることを示している． ALDP のターゲッティングに必要な領域は 67―164 番目のアミノ酸に存在することが報告されてい る．20) このことから,Y174C の変異による構造変化 のため,ターゲッティングシグナルがマスクされて いるのかもしれない．このタイプの変異は ALDP のペルオキシソームへの局在化を調べる上で重要と 考えられる． ALDP の変異で最も多いミスセンス変異ではそ の多くが細胞内で分解を受けている．R617H 及び H667D では発現量の著しい低下が認められる．特 に安定発現した CHO では immunoblot で検出でき なかった．ミスフォールドタンパク質の分解システ ムの 1 つにプロテアソームによる分解系がある．こ のタンパク質分解は,生物の様々な高次機能の制御 や環境ストレスに応答した恒常性の維持(ストレス 応答,タンパク質の品質管理など)に必須な役割を 担っている．しかし,小胞体を経由して合成される 分泌タンパク質や膜タンパク質に比べて,小胞体を 経由しない細胞内タンパク質の品質管理機構はあま り報告されていない．ALDP は遊離のポリソーム から直接ペルオキシソームに輸送されるが,この過 程でどのように R617H, H667D などの変異が認識 され,プロテアソーム系が働いているか興味深い． 171171No.
X
ABCD1 p.Ser606Leu 17202797:53:1304
status: NEW27 df; b9; bb; f3; b9;᜕ᶒఔᢝ௸ ALDP IJe;d30;Pde;ᑁ4d5;ɦb; ߟe00;Έe;ឋ˿a;Ife;IJb;ఐĴb;Ye3;᪆ ALD <a3;ὅIJe;ᢝ௸᜕ᶒ ALDP IJe;a5f;Pfd;,d30;Pde;ᑁc40;ᙠ ឋ,d30;Pde;ᑁIJb;İa;௫Ĵb;b89;b9a;ឋఔYe3;᪆௳Ĵb;௭IJf;, ALDP IJe;ᔜ c9; e1; a4; f3;IJe;a5f;Pfd;ఔMe5;Ĵb;e0a;ᨵᵨIJa;<c5;ᛇ ఔ?d0;f9b;௳Ĵb;əd;Ĵf;Ĵc;Ĵb;&#ff0e;ᱯIJb; df; b9; bb; f3; b9;᜕ᶒIJf;,ıf; ௷ıf; 1 ௸IJe; a2; df; ce;⏚᜕ᶒIJb;ఐĴb;ᶒe38;Ĵb;IJe;ᱯIJb; ‐ᕡdf1;&#ff0e;Ĵf;Ĵc;Ĵf;Ĵc;IJf; ALD <a3;ὅᛇȠa;௯Ĵc;௺Ĵb; df; b9; bb; f3; b9;᜕ᶒIJe;e2d;İb;,TMD İb; 4 ௸(R104C, G116R, Y174C, S342P) ,NBD İb; 4 ௸(Q544R, S606P, S606L, R617H) ,C ʠb;aef;Ze8;f4d;İb; 1 ௸ (H667D)ఔefb;ɢf;IJb;⍶ఁ(Fig. 1) ,ıd;IJe;a5f;Pfd;d30;Pde; ᑁ4d5;ɦb;ఔYe3;᪆௱ıf;&#ff0e;௭Ĵc;IJe;b9f; a13;IJf;,ᜧb66;▾ b7; f3; dd; b8; a6; e0;ᛇȠa;௱ıf;IJe;,a73;௱İf;ff0;ఇıf;əd;௦&#ff0e; ALDP IJf; da; eb; aa; ad; b7; bd; fc; e0;IJb;İa;௫Ĵb;ᬿ╩⒴ᾦPaa; ⏚IJe; b ⏚ᓄIJb;_a2;e0e;௱௺Ĵb;௭İc;Me5;Ĵc;௺Ĵb;&#ff0e; b9f;ωb;IJb; ALD <a3;ὅᵫᩭIJe;e4a;dad;Rbd;d30;Pde;IJf;ᬿ╩⒴ᾦPaa; ⏚IJe; b ⏚ᓄd3b;ឋİc;b63;e38;IJa;dda;dad;Rbd;d30;Pde;bd4;ఇ௺d04; 50 ߟ70%a0b;ea6;e1b;c11;௱௺Ĵb;&#ff0e;ıd;௭[ce;˯f;ɂb;5ca;ఁ᜕ᶒɂb; ALDP IJe;a5f;Pfd;ఔNba;a8d;௳Ĵb;ıf;ఉ,ALDP ఔ˿a;Ife;௱௺ IJa; ALD <a3;ὅᵫᩭdda;dad;Rbd;d30;Pde;IJb;,N ʠb;aef;IJb; His bf; b0;ఔed8;4a0;௱ıf;[ce;˯f;ɂb;᜕ᶒɂb; ALDP ఔe00;Έe;ឋIJb; ˿a;Ife;௱, &#ff3b;1-14 C]lignoceric acid ఔ9fa;cea;௱௺ᬿ╩⒴ ᾦPaa;⏚ b ⏚ᓄd3b;ឋIJe;e2c;b9a;ఔʹc;௷ıf;&#ff0e;ıd;IJe;d50;ʧc;, ALDP b20;ʀd;dda;dad;Rbd;d30;Pde;IJe;ᬿ╩⒴ᾦPaa;⏚ b ⏚ᓄd3b;ឋ IJf;,b63;e38;d30;Pde;IJe;d04; 50%ije;e1b;c11;௱௺ıf;İc;,[ce;˯f; ɂb; His-ALDP ఔ˿a;Ife;௯ıb;Ĵb;b63;e38;Ȝc;a0b;ea6;IJb;ije; d3b;ឋİc;8de;fa9;௱ıf;&#ff0e;௭IJe;௭İb;˿a;Ife;௯ıb;ıf;[ce;˯f;ɂb; His-ALDP IJf; ALDP Ȝc;b49;IJe;a5f;Pfd;ఔᢝ௸௭İc;Nba; a8d;௯Ĵc;ıf;&#ff0e;e00;Ab9;,9 a2e;ϙe;IJe; df; b9; bb; f3; b9;᜕ᶒ ALDP ఔ˿a;Ife;௱ıf;dda;dad;Rbd;d30;Pde;IJf;ᬿ╩⒴ᾦPaa;⏚ b ⏚ᓄd3b; ឋIJe;ᜉ4a0;IJf;a8d;ఉĴc;IJa;İb;௷ıf;&#ff0e;ఐ௷௺,௭Ĵc;IJe; df; b9; bb; f3; b9;᜕ᶒ ALDP IJf;a5f;Pfd;ఔb20;İf;௭İc;Nba;a8d;௯Ĵc; ıf;&#ff0e; ௸,[ce;˯f;ɂb;5ca;ఁ᜕ᶒɂb; His-ALDP ఔ˿a;Ife;௱ ıf; ALD <a3;ὅdda;dad;Rbd;d30;Pde;ఔ8de;5ce;௱,᜕ᶒɂb; ALDP IJe;˿a;Ife;[cf;ఔ immunoblotting IJb;ఐĴa;b9a;[cf;ᓄ௱Ye3;᪆௱ ıf;(Table 1) &#ff0e;IJa;İa; ALDP IJe;˿a;Ife;[cf;IJf;, da; eb; aa; ad; b7; bd; fc; e0;IJe;ᢣa19;⏗d20;Ĵb; ab; bf; e9; fc; bc;IJe;˿a;Ife;[cf;Xdc; b63;௱ıf;&#ff0e;ıd;IJe;d50;ʧc;,᜕ᶒɂb; ALDP(R104C, G116R, Y174C, S342P, Q544R, S606P)IJf;,[ce;˯f;ɂb;ijb;ijc; Ȝc;a0b;ea6;IJe;˿a;Ife;[cf;ఔ̙a;௱ıf;&#ff0e;e00;Ab9;,᜕ᶒɂb; ALDP (S606L, R617H, H667D)IJf;˿a;Ife;[cf;İc;[ce;˯f;ɂb;IJe;˿a; 167 Table 1.
X
ABCD1 p.Ser606Leu 17202797:27:1228
status: NEWX
ABCD1 p.Ser606Leu 17202797:27:5263
status: NEW34 167 No. 1 Ife;[cf;bd4;ఇ௺d04; 50%a0b;ea6;e1b;c11;௱௺ıf;&#ff0e;IJa;İa;,ᔜ ALDP dd; b8; c6; a3; d6;IJe;d30;Pde;IJf;d04; 30%a0b;ea6;Ĵa;,ᔜ d30;Pde;╹IJe;˿a;Ife;4b9;᳛IJb;ᨵɢf;IJa;dee;IJf;a8d;ఉĴc;IJa;İb;௷ ıf;&#ff0e;௭IJe;௭İb;,ALDP IJe;˿a;Ife;[cf;İc;e1b;c11;௱௺ ıf; 3 ௸IJe;᜕ᶒ ALDP IJf;d30;Pde;ᑁIJe;b89;b9a;ឋİc;f4e;e0b;௱ ௺Ĵb;?a8;bdf;௯Ĵc;ıf;&#ff0e;ije;ıf;‐ᕡdf1;௭IJb; S606P S606L IJf;Ȝc;௲Ze8;f4d;IJe;᜕ᶒIJb;ఊfc2;Ĵf;ıa;,f6e;?db;௱ ıf; a2; df; ce;⏚IJb;ఐ௷௺˿a;Ife;[cf;IJb;IJf;dee;İc;a8d;ఉĴc;ıf;&#ff0e; ௸,᜕ᶒɂb; His-ALDP IJe;d30;Pde;ᑁc40;ᙠఔVcd;ᐝ ᢙf53;cd5;Nba;a8d;௱ıf;&#ff0e;᜕ᶒɂb; ALDP(R104C, G116R, S342P, Q544R, S606P, S606L)IJf; ALDP İc; ab; bf; e9; fc; bc;IJe;c40;ᙠe00;Qf4;௱ıf;௭İb;,b63;e38;IJb; da; eb; aa; ad; b7; bd; fc; e0;ఆc40;ᙠ௱௺Ĵb;௭İc;Nba;a8d;௯Ĵc;ıf;&#ff0e;e00;Ab9;, ᜕ᶒɂb; ALDP(Y174C, H667D)IJf;c40;ᙠİc;e00;Qf4;ıb; ıa;,ALDP İc;ed6;IJe;d30;Pde;ᑁc0f;ᘤb98;ఆ╹⍟௷௺f38;〈௯ Ĵc;௺Ĵb;ὃ௨Ĵc;ıf;&#ff0e;᜕ᶒɂb; ALDP(R617H) IJf; ALDP IJe;˿a;Ife;İc;a8d;ఉĴc;IJa;İb;௷ıf;&#ff0e;᜕ᶒɂb; ALDP(R104C, G116R, S342P, Q544R, S606P) IJf;[ce;˯f;ɂb;ijb;ijc;Ȝc;a0b;ea6;IJe; bf; f3; d1; af;[cf;İc;˿a;Ife;௱, da; eb; aa; ad; b7; bd; fc; e0;ఆIJe;c40;ᙠఊNba;a8d;௯Ĵc;ıf;IJe;,௭Ĵc;IJe; ᜕ᶒɂb; ALDP IJf;ᔠᡂ௯Ĵc;ıf;IJe;௵IJb;b63;e38;IJb; da; eb; aa; ad; b7; bd; fc; e0;IJb;Έb;Ĵc;Ĵb;İc;, da; eb; aa; ad; b7; bd; fc; e0;̳c;IJb;İa; ௺ıd;IJe;a5f;Pfd;(ATP d50;ᔠ fb;4a0;c34;ᑖYe3;Re5;௱İf;IJf;9fa;cea;f38; 〈)IJb;ᶒe38;ఔᢝ௸௭İc;?a8;bdf;௯Ĵc;ıf;&#ff0e;ᱯIJb; R104C, G116R, S342P IJf; TMD IJb;b58;ᙠ௳Ĵb;௭İb; ALDP IJe;9fa;cea;f38;〈Pfd;İc;᜕ᓄ௱௺Ĵb;ὃ௨Ĵc;Ĵb;&#ff0e;e00;Ab9;, NBD IJb;b58;ᙠ௳Ĵb; Q544R, S606P IJf; ATP d50;ᔠ fb;4a0;c34; ᑖYe3;IJb;f71;aff;ఔe0e;௨௺Ĵb;5ef;Pfd;ឋİc;ὃ௨Ĵc;Ĵb;&#ff0e;ije;ıf; S606P, S606L IJf;᜕ᶒİc;Ȝc;௲Ze8;f4d;ఊEcb;⌼ḄIJb;b89;b9a; ឋİc;ᶒIJa;௷௺ıf;&#ff0e;Roerig IJf; S606L IJe;᜕ᶒɂb; ALDP IJf;,ATP IJe;Yaa;Ȥc;ឋİc;f4e;e0b;௱௺Ĵb;e00;Ab9; ATP 4a0;c34;ᑖYe3;IJf;b63;e38;IJb;ʹc;Ĵf;Ĵc;௺Ĵb;ᛇȠa;௱௺ Ĵb;&#ff0e; 29) ௭IJe;௭IJf; ALDP ATP IJe;Yaa;Ȥc;ឋİc; ALDP IJe;b89;b9a;ឋIJb;ఊf71;aff;ఔ5ca;ijc;௱௺Ĵb;5ef;Pfd;ឋఔ̙a;௱௺ Ĵb;&#ff0e;S606L S606P IJe;b89;b9a;ឋIJe;⍟a5f;Pfd;IJe;_a2;fc2; IJf; ALDP IJe;a5f;Pfd;ఔMe5;Ĵb;e0a;ఊ‐ᕡdf1;Fb9;Ĵa;, eca;f8c;௯IJb;ʳc;a0e;ఔʹc;௦fc5;⌕İc;Ĵb;&#ff0e;e00;Ab9;,Y174C IJe;᜕ᶒɂb; ALDP IJf;b63;e38;IJb;˿a;Ife;௳Ĵb;IJb;ఊfc2;Ĵf;ıa;, da; eb; aa; ad; b7; bd; fc; e0;ఆc40;ᙠıb;ıa;ed6;IJe;d30;Pde;ᑁc0f;ᘤb98;ఆ df; b9; bf; fc; b2; c3; c6; a3; f3; b0;௱ıf;&#ff0e;௭Ĵc;ije;IJb; da; eb; aa; ad; b7; bd; fc; e0;ఆIJe;c40;ᙠᓄ b7; b0; ca; eb;ఔb20;İf; da; eb; aa; ad; b7; bd; fc; e0; ̳c; bf; f3; d1; af;cea;IJf;,Ϗe;ᱯᶒḄIJb; df; c8; b3; f3; c9; ea; a2;ఌc0f;Pde; f53;IJb;Ofb;ʹc;௳Ĵb;௭İc;Me5;Ĵc;௺Ĵb;&#ff0e; 30,31) ఐ௷௺, ALDP IJe; TMD2ߟ3 IJe;╹IJe; eb; fc; d7;IJf;, da; eb; aa; ad; b7; bd; fc; e0;ఆIJe;c40;ᙠᓄIJb;[cd;⌕IJa;f79;ᒘఔʧc;ıf;௱௺Ĵb;5ef;Pfd; ឋİc;?a8;bdf;௯Ĵc;Ĵb;&#ff0e;Pex19p b58;ᙠᓄIJe; in vitro bf; f3; d1; af;cea;ffb;a33;cfb;IJb;İa;௺,ALDP(Y174C)IJf; Pex19p IJb;d50;ᔠİd;Ĵb;IJe;,ALDP IJe; N ʠb;aef; 67ߟ164 IJb;b58;ᙠ௳Ĵb; da; eb; aa; ad; b7; bd; fc; e0;Ofb;ʹc;IJb;fc2;Ĵf;Ĵb;♚9df;İc; ALDP IJe;f55;İb;IJe;Ecb;⌼᜕ᓄIJb;ఐ௷௺ de; b9; af;௯Ĵc;Ĵb; IJe;İb;ఊ௱Ĵc;IJa;&#ff0e; 5.
X
ABCD1 p.Ser606Leu 17202797:34:844
status: NEWX
ABCD1 p.Ser606Leu 17202797:34:1368
status: NEWX
ABCD1 p.Ser606Leu 17202797:34:3651
status: NEWX
ABCD1 p.Ser606Leu 17202797:34:3855
status: NEW41 168 Vol. 127 (2007) ALDP IJb;௸௺IJf; ALDP IJe; C ʠb;aef;IJb; GFP(green ߮uorescent protein)ఔͮd;ᔠ௯ıb;ıf;᜕ᶒ ALDP-GFP ఔ˿a;Ife;௯ıb;Ĵb;ఊIJb;,[ce;˯f;ɂb; d2; c8; ALDP ఔ CHO d30;Pde;IJb;ᐳ ˿a;Ife;௯ıb;ıf;(CHO d30;Pde;IJb;ఊ ᑁᙠឋIJe; ALDP İc;˿a;Ife;௱௺Ĵb;İc;,ʠc;b9f; a13;IJb;ᵨıf;ᢙf53;İc; ea4;dee;௱IJa;ıf;ఉ, d2; c8; ALDP ఔ˿a;Ife;௯ıb;ıf;) &#ff0e;GFP ͮd;ᔠ bf; f3; d1; af;cea;IJf;ᑁᙠឋIJe; bf; f3; d1; af;cea;IJe;ȕa;ᑩİc;bb9; ᧕Ĵb;௭,b89;b9a;˿a;Ife;d30;Pde;IJe;5d6;f97;İc;bb9;᧕IJb;İd;Ĵb; ௭IJa;IJe;ᑭFb9;İc;Ĵb;&#ff0e;௭IJe;b9f; a13;IJf;, da; eb; aa; ad; b7; bd; fc; e0;IJb;b63;e38;IJb;f38;〈௯Ĵc;Ĵb;᜕ᶒɂb; ALDP(R104C, G116R, S606P) , da; eb; aa; ad; b7; bd; fc; e0;IJb;c40;ᙠ௱IJa;᜕ ᶒɂb; ALDP(Y174C) ,˿a;Ife;[cf;İc;f4e;e0b;௱௺Ĵb;᜕ᶒ ɂb; ALDP(H667D)ఔ⍶క௴&#ff0e;௭Ĵc;b89;b9a;Έe;ᒖ˿a; Ife;d30;Pde;IJb;İa;௫Ĵb; ALDP-GFP IJe;d30;Pde;ᑁc40;ᙠឋఔijf;Ĵb; ,ıd;IJe;ᑖe03;IJf;e00;Έe;ឋ˿a;Ife;௯ıb;ıf; His-ALDP Ȝc; Ed8;௷ıf;&#ff0e; f97;Ĵc;ıf;b89;b9a;˿a;Ife;d30;Pde;ఐĴa; aa; eb; ac; cd; e9;c97;ᑖ˱b;ఔabf; Xfd;௱, b7; e7;cd6;bc6;ea6;4fe;Βd;⍤fc3;ᑖ`e2;cd5;IJb;ఐĴa;ᔜ d5; e9; af; b7; e7; f3;IJb;ᑖ௫ıf;IJe;௵,SDS-PAGE 5ca;ఁ immunoblotting IJb;ఐĴa;᜕ᶒ ALDP IJe;c40;ᙠIJb;௸௺Ye3;᪆ఔʹc;௷ ıf;(Fig. 3) &#ff0e; da; eb; aa; ad; b7; bd; fc; e0; de; fc; ab; fc;௱௺ da; eb; aa; ad; b7; bd; fc; e0;̳c; bf; f3; d1; af;cea;Ĵb; PMP70 da; eb; aa; ad; b7; bd; fc; e0;IJe;e3b;⌕IJa; de; c8; ea; c3; af; b9; bf; f3; d1; af;cea; Ĵb; ab; bf; e9; fc; bc;ఔᵨıf;&#ff0e;[ce;˯f;ɂb; ALDP ఔb89;b9a;Έe;ᒖ ˿a;Ife;௱௺Ĵb;d30;Pde;IJb;İa;௺, ab; bf; e9; fc; bc;d3b;ឋe26;ఁIJb; PMP70 İc;e3b;௱௺ d5; e9; af; b7; e7; f3; 3 5ca;ఁ 4 IJb;b58;ᙠ௳ Ĵb;௭ఐĴa;,௭IJe;ᑖ˱b;IJb; da; eb; aa; ad; b7; bd; fc; e0;İc;8de;5ce;௯ Ĵc;ıf;௭İc;̙a;ᖂ௯Ĵc;ıf;&#ff0e;ije;ıf;d04; 110 kDa IJe;ᑖb50; b5; a4; ba;ఔᢝ௸ ALDP-GFP e26;ఁIJb; 83 kDa IJe;[ce;˯f;ɂb; ALDP IJf;, da; eb; aa; ad; b7; bd; fc; e0; de; fc; ab; fc;ijb;ijc;Ȝc;௲ ᑖe03;ఔ̙a;௱௺ıf;௭İb; da; eb; aa; ad; b7; bd; fc; e0;IJb;c40;ᙠ ௱௺Ĵb;௭İc;̙a;ᖂ௯Ĵc;ıf;&#ff0e;ije;ıf;᜕ᶒɂb; ALDP-GFP(G116R, S606P)ఊȜc;Ed8;IJe;ᑖe03;ఔ̙a;௱ıf;&#ff0e; e00;Ab9;,᜕ᶒɂb; ALDP-GFP(H667D)ఔb89;b9a;Έe;ᒖ ˿a;Ife;௱௺Ĵb;d30;Pde;IJe;ᛊᔠIJf;,ALDP-GFP IJe; d0; f3; c9; IJf;ʳc;3fa;௯Ĵc;IJa;İb;௷ıf;(Fig. 3) &#ff0e;‐ᕡdf1;௭IJb;, PMP70 IJf; ʳc; 3fa; ௯ Ĵc; ıf; İc; , ᐳ ˿a; Ife; ௯ ıb; ıf; [ce; ˯f; ɂb; ALDP IJe; d0; f3; c9;ఊʳc;3fa;௯Ĵc;IJa;İb;௷ıf;&#ff0e;ije;ıf;᜕ᶒɂb; ALDP(S606L)IJb;௸௺ఊȜc;Ed8;௷ıf;&#ff0e;௭Ĵc; IJe; d50; ʧc; IJf; , ᜕ ᶒ ɂb; ALDP ( H667D, S606L ) IJf; PMP70 IJf;IJa;İf;,[ce;˯f;ɂb; ALDP ⋋ᔠf53;ఔf62;ᡂ ௱,e21;ὅİc;ᑖYe3;௯Ĵc;Ĵb;5ef;Pfd;ឋఔ̙a;ᖂ௱௺Ĵb;&#ff0e; ABC bf; f3; d1; af;cea;IJe;a5f;Pfd;˿a;Ife;IJb;[cd;⌕Ĵb; TMD ఌ NBD ee5;IJe; C ʠb;aef;Ze8;f4d;IJe;᜕ᶒİc; bf; f3; d1; af;cea;IJe;b89; b9a;ឋIJb;f71;aff;ఔ5ca;ijc;௳௭IJf;‐ᕡdf1;&#ff0e;ALDP IJe; C ʠb;aef;Ze8;f4d;Ĵb; 600ߟ700 a2; df; ce;⏚IJe;᜕ᶒİc; X-ALD ఔf15;İd;d77;௭௳ϗb;ea6;İc; ad8;௭İb;,ALDP IJe; C ʠb;aef;Ze8;f4d;IJf; bf; f3; d1; af;cea;IJe;b89;b9a;ឋIJb;[cd;⌕IJa;f79;ᒘఔ>c5; ௷௺Ĵb;5ef;Pfd;ឋİc;Ĵb;&#ff0e;Liu IJf; ALDP IJe; c0; a4; de; fc;ᓄIJb;IJf; C ʠb;aef;Ze8;f4d;(AA.631ߟ745)İc;[cd;⌕ Ĵb;ᛇȠa;௱௺Ĵb;&#ff0e; 13) H667D ఌ S606L IJe;ఐ௦IJa; ᜕ᶒIJf;,ıd;Ĵc;Qea;eab;Ĵb;IJf;[ce;˯f;ɂb; ALDP df; b9; d5; a9; fc; eb; c9;௱ıf; c0; a4; de; fc;ఔf62;ᡂ௱,ᶒe38; bf; f3; d1; af;cea; ௱௺a8d;b58;௯Ĵc;ᑖYe3;௯Ĵc;Ĵb;ὃ௨Ĵc;Ĵb;&#ff0e;e00;Ab9;,᜕ᶒ ɂb; ALDP-GFP(R104C)IJf; da; eb; aa; ad; b7; bd; fc; e0;ᑖ˱b; IJb;8de;5ce;௯Ĵc;Ĵb;ఊIJe;IJe;, d5; e9; b0; e1; f3; c8;ᓄ௱௺Ĵb;௭ 169 Fig. 4.
X
ABCD1 p.Ser606Leu 17202797:41:4880
status: NEWX
ABCD1 p.Ser606Leu 17202797:41:5064
status: NEWX
ABCD1 p.Ser606Leu 17202797:41:6480
status: NEW46 ᜕ᶒɂb; ALDP IJe;ᑖYe3;Έe;a0b;IJe;Ye3;᪆ Ab0;˯f; bf; f3; d1; af;cea;İc;b63;௱ d5; a9; fc; eb; c7; a3; f3; b0;ఔ5d7;௫ Ĵb;௭IJf;,ıd;IJe; bf; f3; d1; af;cea;IJe;b63;e38;IJa;a5f;Pfd;˿a;Ife;IJe;ıf;ఉ IJb;fc5;♐Ĵb;&#ff0e;a;f1d;b50;᜕ᶒIJa;İc;b58;ᙠ௳Ĵb;, bf; f3; d1; af;cea;İc; df; b9; d5; a9; fc; eb; c7; a3; f3; b0;௯Ĵc;Ĵb;&#ff0e;௭IJe; df; b9; d5; a9; fc; eb; c9; bf; f3; d1; af;İc;d30;Pde;ఆᑖccc;௯Ĵc;ıf;Ĵa;,d30;Pde;ᑁ IJb;Tc4;a4d;௱ıf;Ĵa;௳Ĵb;˯f;f53;IJb;௷௺ᬿఉ௺ᨵbb3;IJb;IJa;Ĵb; ıf;ఉ,௭IJe;ఐ௦IJa; bf; f3; d1; af;IJf; d7; ed; c6; a2; bd; fc; e0;, ea; bd; bd; fc; e0;b49;IJb;ఐ௷௺fc5;΅f;IJb;ᑖYe3;௯Ĵc;Ĵb;&#ff0e;௵IJa;ijf;IJb;,8a2; Pde;ឋdda;dad;Kc7;IJe;țf;8e0; bf; f3; d1; af;cea; CFTR IJf;d30;Pde;̳c; a4; aa; f3; c1; e3; cd; eb;௱௺a5f;Pfd;௳Ĵb; ABC bf; f3; d1; af;cea;Ĵb; İc;,᜕ᶒ CFTR IJf;c0f;Pde;f53;̳c;İb; d7; ed; c6; a2; bd; fc; e0;IJb; ea; af; eb; fc; c8;௯Ĵc;ᑖYe3;௯Ĵc;Ĵb;௭İc;ᛇȠa;௯Ĵc;௺ Ĵb;&#ff0e; 32,33) ௱İb;௱IJa;İc;,᜕ᶒɂb; ALDP ఔ;cb;ఉ௱ ௺, da; eb; aa; ad; b7; bd; fc; e0;̳c; bf; f3; d1; af;cea;IJb;௸௺IJe;Ye3;᪆ IJf;ijb;కʹc;Ĵf;Ĵc;௺IJa;&#ff0e; ᜕ᶒɂb; ALDP IJe;e00;Έe;ឋ˿a;Ife;b89;b9a;Έe;ᒖ˿a;Ife;b9f; a13; ఐĴa;,ALDP (S606L, R617H, H667D, R104C) IJf;, d7; ed; c6; a2; fc; bc;IJb;ఐĴa;ᑖYe3;௯Ĵc;௺Ĵb;?a8;b9a;௯Ĵc;ıf;&#ff0e; ıd;௭,ALDP-GFP(H667D)ఔ˿a;Ife;௱௺Ĵb; CHO d30;Pde;IJb;ᔜa2e; d7; ed; c6; a2; fc; bc;σb;bb3;ᒐఔ3e6;ᳮ௱,Ye3; ᪆ఔʹc;௷ıf;&#ff0e;ıd;IJe;d50;ʧc;, d7; ed; c6; a2; bd; fc; e0;σb;bb3;ᒐ Ĵb; lactacystin ఔ3e6;ᳮ௱ıf;d30;Pde;IJf; ALDP-GFP 5ca;ఁ ALDP IJe; d0; f3; c9; İc; 3fa; Ife; ௱ ıf; ( Fig. 4 ) &#ff0e; e00; Ab9; , leupeptin, AEBSF, E64d IJb;IJf;4b9;ʧc;İc;IJa;İb;௷ıf;&#ff0e;ije; ıf;ed6;IJe; d7; ed; c6; a2; bd; fc; e0;σb;bb3;ᒐĴb; MG132 ఊᨵ4b9; ௷ıf;&#ff0e;௯IJb; d7; ed; c6; a2; bd; fc; e0;σb;bb3;ᒐIJb;ఐĴa;ᑖYe3; ఔ⌫Ĵc;ıf;᜕ᶒɂb; ALDP-GFP(H667D)IJe;d30;Pde;ᑁc40; ᙠఔVcd;ᐝᢙf53;cd5;Yb3;bdf;௳Ĵb;, da; eb; aa; ad; b7; bd; fc; e0;IJb; c40;ᙠ௱௺Ĵb;௭İc;Nba;a8d;௯Ĵc;ıf;&#ff0e;e00;Ab9;,᜕ᶒɂb; ALDP (R104C) IJe; d5; e9; b0; e1; f3; c8;ᓄIJf;e0a;a18; d7; ed; c6; a2; fc; bc;3e6;ᳮIJf;σb;bb3;௯Ĵc;IJa;İb;௷ıf;&#ff0e; ௯IJb; ALD <a3;ὅᵫᩭd30;Pde;IJe;ᑁ8e0;ឋ᜕ᶒ ALDP IJe;ᑖYe3; d7; ed; c6; a2; bd; fc; e0;ᑖYe3;cfb;IJe;_a2;e0e;IJb;௸௺Nba;a8d; ௳Ĵb;ıf;ఉ,᜕ᶒɂb; ALDP(R617H)ఔᢝ௸<a3;ὅᵫ ᩭdda;dad;Rbd;d30;Pde;ఔᵨ௺ bf; f3; d1; af;ᑖYe3;IJe;σb;bb3;b9f; a13;ఔʹc; ௷ıf;&#ff0e;ıd;IJe;d50;ʧc;,lactacystin MG132 3e6;ᳮIJb;ఐĴa;, ALDP IJe; d0; f3; c9;İc;3fa;Ife;௱ıf;&#ff0e;ee5;e0a;IJe;d50;ʧc;ఐĴa;, da; eb; aa; ad; b7; bd; fc; e0;̳c;e0a;IJb;IJf; df; b9; d5; a9; fc; eb; c9;௱ıf; bf; f3; d1; af;cea;ఔa8d;b58;௳Ĵb;ed5;d44;ijf;İc;b58;ᙠ௱, d7; ed; c6; a2; bd; fc; e0;5ca; ఁed6;IJe; d7; ed; c6; a2; fc; bc;ఔecb;௱௺◀௱௺Ĵb;௭İc;̙a; ᖂ௯Ĵc;ıf;&#ff0e; e00;Ab9;,c71;ᵪIJf; ALD <a3;ὅdda;dad;Rbd;d30;Pde;ఔ &#ff3b;35 S&#ff3d; e1; c1; aa; cb; f3; d1; eb; b9; c1; a7; a4; b9;௳Ĵb;௭IJb;ఐĴa;,᜕ᶒɂb; ALDP (G512S, R660W) IJe;ᑖYe3;İc; E-64 leupepu- tin IJb;ఐĴa;ᢓᑴ௯Ĵc;Ĵb;௭ఔᛇȠa;௱௺Ĵb;&#ff0e; 34) f7c; IJe;b9f; a13;IJf; d7; ed; c6; a2; bd; fc; e0;σb;bb3;ᒐIJb;௸௺IJf;b9f; a13;௱ ௺IJa;IJe;, d7; ed; c6; a2; bd; fc; e0;IJe;_a2;e0e;IJf;e0d;ʔe;Ĵb; İc;,᜕ᶒɂb; ALDP IJe;ᑖYe3;IJb;IJf;,⋋ᦪIJe; d7; ed; c6; a2; fc; bc;İc;_a2;e0e;௱௺Ĵb;5ef;Pfd;ឋİc;Ĵb;&#ff0e; 7.
X
ABCD1 p.Ser606Leu 17202797:46:2435
status: NEW
PMID: 22479560
[PubMed]
Pereira Fdos S et al: "Mutations, clinical findings and survival estimates in South American patients with X-linked adrenoleukodystrophy."
No.
Sentence
Comment
24
Family/Index case Phenotype at diagnosis Mutation Exon/IVS Mutation type Effect on protein (cDNA) Effect on protein (mRNA) Protein localization Origin of mutations Origin of family 1/Female asymptomatic p.Gly512Ser (Feigenbaum V et al. 1996) E6 Missense c.1534G.A GGC.AGC NBF de novo Southern Brazil 2/Female asymptomatic p.Ser606Leu (Fanen P et al., 1994) E8 Missense c.1817C.T UCG.UUG NBF Inherited Southern Brazil 3/Male AMN p.Trp601X (Gartner J et al.,1998) E8 Stop codon c.1802C.A Truncated NBF Inherited Southern Brazil 4/Female asymptomatic p.Arg617His (Fanen P et al., 1994) E8 Missense c.1850G.A CGC.CAC NBF ND Southern Brazil 5/Male AMN p.Pro623Leu # E9 Missense c.1868C.T CCC.CUC NBF Inherited Southern Brazil 6/Male AO p.Trp326X (Barcelo A et al, 1996) E2 Stop codon c.978G.A Truncated TMD Inherited Southern Brazil 8/Female asymptomatic p.Glu577X # E7 Stop codon c.1729G.T Truncated NBF Inherited Southern Brazil 9/Male asymptomatic p.Arg554His (Smith KD et al., 1999) E7 Missense c.1661G.A CGU.CAU NBF Inherited Southern Brazil 10/Male CALD p.Arg518Gln (Imamura A et al., 1997) E6 Missense c.1553G.A CGG.CAG NBF Inherited Southern Brazil 11/Male AO p.Tyr33_Pro34fsX34 # E1A Frameshift+stop codon c.99_102delC Truncated - Inherited Southern Brazil 12/Female asymptomatic p.Gly266Arg (Fuchs S et al., 1994) E7 Missense c.1653insG Truncated TMD ND Southern Brazil 20/Male CALD p.Arg538fs # E6 Frameshift c.1614_1615dup27 Elonged NBF de novo Southern Brazil 21/Male CALD p.Ala232fsX64 # E2 Frameshift+stop codon c.696_697del11 Truncated TMD Inherited Southern Brazil 22/Male CALD p.Trp137fsX57 # E1B Frameshift+stop codon c.411_412insC Truncated TMD Inherited Northern Brazil 23/Male asymptomatic p.Trp679X (Waterham HR et al, 1998) E10 Stop codon c.2037G.A Truncated NBF ND Southern Brazil 24/Male AO p.Tyr296Cys (Takano H et al., 1999) E2 Missense c.887A.G UAU.UGU TMD Inherited Southern Brazil 27/Male CALD p.Leu628Glu # E9 Missense c.1883T.A CUG.GAG NBF Inherited Southern Brazil 29/Male CALD p.Pro546fsX?
X
ABCD1 p.Ser606Leu 22479560:24:324
status: NEW