ABCD1 p.Ser606Pro
| ClinVar: |
c.1818G>A
,
p.Ser606=
?
, Uncertain significance
c.1817C>T , p.Ser606Leu D , Pathogenic |
| Predicted by SNAP2: | A: D (91%), C: D (95%), D: D (95%), E: D (95%), F: D (95%), G: D (95%), H: D (95%), I: D (95%), K: D (95%), L: D (95%), M: D (95%), N: D (95%), P: D (95%), Q: D (95%), R: D (95%), T: D (95%), V: D (95%), W: D (95%), Y: D (95%), |
| Predicted by PROVEAN: | A: N, C: D, D: D, E: D, F: D, G: D, H: D, I: D, K: D, L: D, M: D, N: N, P: D, Q: D, R: D, T: N, V: D, W: D, Y: D, |
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[hide] Mutational and protein analysis of patients and he... Am J Hum Genet. 1996 Jun;58(6):1135-44. Feigenbaum V, Lombard-Platet G, Guidoux S, Sarde CO, Mandel JL, Aubourg P
Mutational and protein analysis of patients and heterozygous women with X-linked adrenoleukodystrophy.
Am J Hum Genet. 1996 Jun;58(6):1135-44., [PMID:8651290]
Abstract [show]
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.
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No. Sentence Comment
76 58:1135-1144, 1996 Table 2 Mutations in the ALD Gene in Studied Patients AMINO ACID MUTATIONSb HOMOLOGUE INd KINDRED CLINICAL LOCALIZATION AMINO ACID ALDP BY NUMBER PHENOTYPEa DNA CpG Exon IN PROTEINC ALTERATION h/m ALDRP hPMP70 IF/WB' CALD, AMN CALD CALD CALD, AS AD CALD, AMN CALC AD AD AD ALMD CALD CALD, AMN CALD CALD, AMN, AD AMN ALMD CALD ALMD CALD AMN ALD AD, AMN, AS CALD, AS CALD CALD AD CALD AMN, ALMD CALD CALD AMN, ALMD CALD CALD, AMN, ALMD CALD CALD, ALMD, AS ALMD CALD AMN CALD, AMN AD AD AMN CALD G416A Ins T524 C679T C679T C700T C709G G732A A829G C840T Del TA927-28 A928G A985T A1048G DeIGC1080-81 C1174T G1266A ins C1521 1636delC DelAG 1801-02 DelAG 1801-02 DelAG 1801-02 DelAG 1801-02 ins TGG 1848 G 1920 A C1938T C1938T G1950A C2065T C2065T C2065T C2065T C2065G G 2166+1 A T2202C DelGC 2335 C2364T C2364T No mutation found No mutation found No mutation found No mutation found No mutation found No mutation found No mutation found 1 1 + 1 + 1 1 1 + 1 1 + 1 1 1 1 1 1 1 + 1 3 4 S 5 S S S 6 + 6 + 6 6 + 7 + 7 + 7 + 7 + 7 + 7 8 9 9 9 W10 X Frameshift at L46 TMS2 S98L TMS2 S98L T1OSI S108W G116R TMS3 N148S TMS3 R152C Frameshift at Y180 Y181C TMS4 D200V TMS4 D221G Frameshift at R231 P263L EAA-like A294T Frameshift at V378 Frameshift at T416 Frameshift at E471 Frameshift at E471 Frameshift at E471 Frameshift at E471 ins val 491 Walker A G512S Walker A R518W Walker A R518W G 522 W P560L P560L P560L P560L P56OR Splice at G593 Walker B S606P Frameshift at D649 R660W R660W Absent Not done S A Present S A Present T T Absent S D Decreased G T Absent N N Present R K Present Absent Y Y Not done D D Not done D D Absent Absent P R Decreased A A Not done Absent Absent Absent Absent Absent Absent Absent G G Absent R R Absent R R Decreased G E Absent P P Decreased P P Decreased P P Decreased P P Absent P P Absent Not done S S Absent Absent R R Absent R R Absent Not done Absent Absent Absent Present Absent Absent a CALD = cerebral ALD (5-15 years); AMN = adrenomyeloneuropathy; ALMD = adrenomyeloneuropathy with cerebral involvement; AD = Addison disease; AS = Asymptomatic.
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ABCD1 p.Ser606Pro 8651290:76:1454
status: NEW131 Lane 1, protein markers; lane 2, control; lane 3, patient 18 (S108W); lane 4, patient 32 (P263L); lane 5, patient 5 (P560L); lane 6, patient 4 (G116R); lane 7, patient 19 (D221G); lane 8, patient 33 (S98L); lane 9, patient 78 (S606P); lane 10, patient 3 (no mutation found); lane 11, patient 37 (P560L); lane 12, patient 22 (R660W); lane 13, control; lane 14, patient 39 (T1051); lane 15, patient 4 (G116R); lane 16, patient 43 (frameshift at Y180); lane 17, patient 5 (P560L); lane 18, patient 59 (G512S); lane 19, patient 29 (frameshift at D649); lane 20, patient 69 (P560L); lane 21, patient 19 (D221G); lane 22, patient 64 (W1OX); lane 23, patient 63 (frameshift at R231); lane 24, patient 52 (no mutation found); lane 25, patient 61 (frameshift at E471); and lane 26, patient 83 (G522W).
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ABCD1 p.Ser606Pro 8651290:131:227
status: NEW144 Four missense mutations (G512S, R518W, G522W, and S606P) were found in the ATP-binding domain.
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ABCD1 p.Ser606Pro 8651290:144:50
status: NEW145 Three of these mutations (G512S, G522W, and S606P) affect amino acid residues that are identical among other ABC transporters (see fig. 4 in Mosser et al. 1993 and fig. 1 in Fanen et al. 1994).
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ABCD1 p.Ser606Pro 8651290:145:44
status: NEW172 Missense mutations leading to a lack of ALDP included three mutations located in the ATP-binding domain (G512S, G522W, and S606P).
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ABCD1 p.Ser606Pro 8651290:172:123
status: NEW173 Four missense mutations (S108W, P263L, R518W, and P560L) resulted in decreased ALDP immunoreactivity reflecting likely instability and/or partial deficiency in the peroxisomal targeting of ALDP.
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ABCD1 p.Ser606Pro 8651290:173:123
status: NEW75 58:1135-1144, 1996 Table 2 Mutations in the ALD Gene in Studied Patients AMINO ACID MUTATIONSb HOMOLOGUE INd KINDRED CLINICAL LOCALIZATION AMINO ACID ALDP BY NUMBER PHENOTYPEa DNA CpG Exon IN PROTEINC ALTERATION h/m ALDRP hPMP70 IF/WB' CALD, AMN CALD CALD CALD, AS AD CALD, AMN CALC AD AD AD ALMD CALD CALD, AMN CALD CALD, AMN, AD AMN ALMD CALD ALMD CALD AMN ALD AD, AMN, AS CALD, AS CALD CALD AD CALD AMN, ALMD CALD CALD AMN, ALMD CALD CALD, AMN, ALMD CALD CALD, ALMD, AS ALMD CALD AMN CALD, AMN AD AD AMN CALD G416A Ins T524 C679T C679T C700T C709G G732A A829G C840T Del TA927-28 A928G A985T A1048G DeIGC1080-81 C1174T G1266A ins C1521 1636delC DelAG 1801-02 DelAG 1801-02 DelAG 1801-02 DelAG 1801-02 ins TGG 1848 G 1920 A C1938T C1938T G1950A C2065T C2065T C2065T C2065T C2065G G 2166+1 A T2202C DelGC 2335 C2364T C2364T No mutation found No mutation found No mutation found No mutation found No mutation found No mutation found No mutation found 1 1 + 1 + 1 1 1 + 1 1 + 1 1 1 1 1 1 1 + 1 3 4 S 5 S S S 6 + 6 + 6 6 + 7 + 7 + 7 + 7 + 7 + 7 8 9 9 9 W10 X Frameshift at L46 TMS2 S98L TMS2 S98L T1OSI S108W G116R TMS3 N148S TMS3 R152C Frameshift at Y180 Y181C TMS4 D200V TMS4 D221G Frameshift at R231 P263L EAA-like A294T Frameshift at V378 Frameshift at T416 Frameshift at E471 Frameshift at E471 Frameshift at E471 Frameshift at E471 ins val 491 Walker A G512S Walker A R518W Walker A R518W G 522 W P560L P560L P560L P560L P56OR Splice at G593 Walker B S606P Frameshift at D649 R660W R660W Absent Not done S A Present S A Present T T Absent S D Decreased G T Absent N N Present R K Present Absent Y Y Not done D D Not done D D Absent Absent P R Decreased A A Not done Absent Absent Absent Absent Absent Absent Absent G G Absent R R Absent R R Decreased G E Absent P P Decreased P P Decreased P P Decreased P P Absent P P Absent Not done S S Absent Absent R R Absent R R Absent Not done Absent Absent Absent Present Absent Absent a CALD = cerebral ALD (5-15 years); AMN = adrenomyeloneuropathy; ALMD = adrenomyeloneuropathy with cerebral involvement; AD = Addison disease; AS = Asymptomatic.
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ABCD1 p.Ser606Pro 8651290:75:1454
status: NEW[hide] Adrenoleukodystrophy: subcellular localization and... J Neurochem. 2007 Jun;101(6):1632-43. Takahashi N, Morita M, Maeda T, Harayama Y, Shimozawa N, Suzuki Y, Furuya H, Sato R, Kashiwayama Y, Imanaka T
Adrenoleukodystrophy: subcellular localization and degradation of adrenoleukodystrophy protein (ALDP/ABCD1) with naturally occurring missense mutations.
J Neurochem. 2007 Jun;101(6):1632-43., [PMID:17542813]
Abstract [show]
Mutation in the X-chromosomal adrenoleukodystrophy gene (ALD; ABCD1) leads to X-linked adrenoleukodystrophy (X-ALD), a severe neurodegenerative disorder. The encoded adrenoleukodystrophy protein (ALDP/ABCD1) is a half-size peroxisomal ATP-binding cassette protein of 745 amino acids in humans. 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. 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. Furthermore, mutant ALDP-green fluorescence proteins (S606L and H667D) stably expressed in CHO cells were not detected due to rapid degradation. Interestingly, the wild type ALDP co-expressed in these cells also disappeared. In the case of X-ALD fibroblasts from an ALD patient (R617H), the mutant ALDP was not detected in the cells, but appeared upon incubation with a proteasome inhibitor. When CHO cells expressing mutant ALDP-green fluorescence protein (H667D) were cultured in the presence of a proteasome inhibitor, both the mutant and wild type ALDP reappeared. In addition, mutant His-ALDP (Y174C), which has a mutation between transmembrane domain 2 and 3, did not exhibit peroxisomal localization by immunofluorescense study. 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. Further, the region between transmembrane domain 2 and 3 is important for the targeting of ALDP to the peroxisome.
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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.Ser606Pro 17542813:2:99
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.Ser606Pro 17542813:35:160
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.Ser606Pro 17542813:71:479
status: NEW119 Six mutant His-ALDPs (R104C, G116R, Y174C, S342P, Q544R, and S606P) were expressed in an equal amount to the wild type His-ALDP.
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ABCD1 p.Ser606Pro 17542813:119:62
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.Ser606Pro 17542813:127:60
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.Ser606Pro 17542813:135:47
status: NEW139 Wild type ALDP-GFP and mutant ALDP-GFP (G116R and S606P) were recovered mainly in the fractions 3 and 4 and the distribution was similar to those of PMP70 and catalase, suggesting that these ALDP-GFPs had located to peroxisomes as had the corresponding His-ALDPs in 163T cells.
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ABCD1 p.Ser606Pro 17542813:139:50
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.Ser606Pro 17542813:150:178
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.Ser606Pro 17542813:180:274
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.Ser606Pro 17542813:181:151
status: NEW217 S606P and S606L are in the ABC signature motif.
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ABCD1 p.Ser606Pro 17542813:217:0
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.Ser606Pro 17542813:229:343
status: NEW259 Second, mutant ALDP (G116R, S342P, Q544R, and S606P) expressed similar levels to wild type ALDP in the experiment of transient expression as the corresponding His-ALDP in X-ALD fibroblasts (Fig. 3b and Table 2) and stable expression as ALDP-GFP in CHO cells (Fig. 4 and Table 2).
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ABCD1 p.Ser606Pro 17542813:259:46
status: NEW263 The substitution of Q554R and S606P located to NBD, which might affect the binding and hydrolysis of ATP in ALDP.
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ABCD1 p.Ser606Pro 17542813:263:30
status: NEW[hide] X-linked adrenoleukodystrophy: ABCD1 de novo mutat... Mol Genet Metab. 2011 Sep-Oct;104(1-2):160-6. Epub 2011 Jun 22. Wang Y, Busin R, Reeves C, Bezman L, Raymond G, Toomer CJ, Watkins PA, Snowden A, Moser A, Naidu S, Bibat G, Hewson S, Tam K, Clarke JT, Charnas L, Stetten G, Karczeski B, Cutting G, Steinberg S
X-linked adrenoleukodystrophy: ABCD1 de novo mutations and mosaicism.
Mol Genet Metab. 2011 Sep-Oct;104(1-2):160-6. Epub 2011 Jun 22., [PMID:21700483]
Abstract [show]
X-linked adrenoleukodystrophy (X-ALD) is a progressive peroxisomal disorder affecting adrenal glands, testes and myelin stability that is caused by mutations in the ABCD1 (NM_000033) gene. Males with X-ALD may be diagnosed by the demonstration of elevated very long chain fatty acid (VLCFA) levels in plasma. In contrast, only 80% of female carriers have elevated plasma VLCFA; therefore targeted mutation analysis is the most effective means for carrier detection. Amongst 489 X-ALD families tested at Kennedy Krieger Institute, we identified 20 cases in which the ABCD1 mutation was de novo in the index case, indicating that the mutation arose in the maternal germ line and supporting a new mutation rate of at least 4.1% for this group. In addition, we identified 10 cases in which a de novo mutation arose in the mother or the grandmother of the index case. In two of these cases studies indicated that the mothers were low level gonosomal mosaics. In a third case biochemical, molecular and pedigree analysis indicated the mother was a gonadal mosaic. To the best of our knowledge mosaicism has not been previously reported in X-ALD. In addition, we identified one pedigree in which the maternal grandfather was mosaic for the familial ABCD1 mutation. Less than 1% of our patient population had evidence of gonadal or gonosomal mosaicism, suggesting it is a rare occurrence for this gene and its associated disorders. However, the residual maternal risk for having additional ovum carrying the mutant allele identified in an index case that appears to have a de novo mutation is at least 13%.
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No. Sentence Comment
90 Previously reported (Yes/No) Number of pedigrees reporteda CpG (Yes/No) 25 c.124delC ND No 1 N/A 5 c.279_280ins12bp (p.Leu93_Leu94insGluThrGlyLeu) ND No 1 No 6 c.410G>A (p.Trp137X) ND No 1 No 7 c.412_414delCTC (p.Leu139del) ND Yes 2 No 26 c.476del24 ND No 1 N/A 4 c.593C>G (p.Thr198Met) N/A No 1 Yes 8 c.695_696insG (p.Ala233fsX67) ND No 1 Yes 2 c.725G>A (p.Trp242X) Gonosomal No 1 No c.796G>A (p.Gly266Arg) ND Yes 23 Yes 27 c.797G>A (p.Gly266Gln) ND No 1 No 12 c.944C>A (p.Ser315X) ND No 1 Yes 13 c.1201C>T (p.Arg401Trp) ND Yes 12 Yes 14 c.1225-2A>G (splice defect) ND No 1 No 15 c.1390C>T (p.Arg464X) ND Yes 11 Yes 16 c.1553G>A (p.Arg518 Gln) ND Yes 20 Yes 17 c.1567C>T (p.Leu523Phe) ND No 1 No 18 c.1609C>T (p.Gln537X) ND No 1 No 28 c.1619T>G (p.Phe540Cys) ND No 1 No 19 c.1679C>T (p.Pro560Leu) ND Yes 20 Yes 29 c.1679C>T (p.Pro560Leu) ND Yes 20 Yes 20 exon3 to exon10 deletion ND Yesb 9 N/A 30 c.1781-1G>A ND No 1 No 21 c.1816T>C (p.Ser606Pro) ND Yes 3 No 31 c.1850G>A (p.Arg617His) ND Yes 20 Yes 22 c.1876G>A (p.Ala626Thr) ND Yes 10 Yes 23 c.1894A>C (p.Thr632Pro) ND No 2 No 1 c.1899C>A (p.Ser633Arg) Gonosomal Yes 2 Yes 24 c.1918 G>A (p.Glu640Lys) ND No 2 No 3 c.2030G>A (p.Gly677Asp) Gonadal No 1 Yes de novo mutation in male index case with childhood cerebral X-ALD;somatic and/or gonadal mosaicisim; de novo mutationND = none detected; N/A = not applicable; Color codes: in female carrier.
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ABCD1 p.Ser606Pro 21700483:90:937
status: NEW[hide] Genomic profiling identifies novel mutations and S... PLoS One. 2011;6(9):e25094. Epub 2011 Sep 22. Kumar N, Taneja KK, Kalra V, Behari M, Aneja S, Bansal SK
Genomic profiling identifies novel mutations and SNPs in ABCD1 gene: a molecular, biochemical and clinical analysis of X-ALD cases in India.
PLoS One. 2011;6(9):e25094. Epub 2011 Sep 22., [PMID:21966424]
Abstract [show]
X-linked adrenoleukodystrophy (X-ALD) affects the nervous system white matter and adrenal cortex secondary to mutations in the ABCD1 gene that encode the peroxisomal membrane protein. We conducted a genomic and protein expression study of susceptibility gene with its clinical and biochemical analysis. To the best of our knowledge this is the first preliminary comprehensive study in Indian population that identified novel mutations and SNPs in a relatively large group. We screened 17 Indian indigenous X-linked adrenoleukodystrophy cases and 70 controls for mutations and SNPs in the exonic regions (including flanking regions) of ABCD1 gene by direct sequencing with ABI automated sequencer along with Western blot analysis of its endogenous protein, ALDP, levels in peripheral blood mononuclear cells. Single germ line mutation was identified in each index case in ABCD1 gene. We detected 4 novel mutations (2 missense and 2 deletion/insertion) and 3 novel single nucleotide polymorphisms. We observed a variable protein expression in different patients. These findings were further extended to biochemical and clinical observations as it occurs with great clinical expression variability. This is the first major study in this population that presents a different molecular genetic spectrum as compared to Caucasian population due to geographical distributions of ethnicity of patients. It enhances our knowledge of the causative mutations of X-ALD that grants holistic base to develop effective medicine against X-ALD.
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No. Sentence Comment
103 Of the 10 non-recurrent mutations in our study group i.e a frameshift mutation c.110_117del8 (Val36fs) in exon 1 was identified in an asymptomatic heterozygous female and 5 missense mutations, c.904G.A (Glu302Lys) in exon 2, c.1202G.A (Arg401Gln) in exon 3, c.1771C.T (Arg591Trp) in exon 7, c.1816T.C (Ser606Pro) and c.1825G.A (Glu609Lys) in exon 8 were present in 5 different patients.
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ABCD1 p.Ser606Pro 21966424:103:302
status: NEW156 Patients Phenotype1 Age(Year) Sex Exon/IVS Mutation Type Mutations Protein Localization ALDP PSIC Score5 P01* ccALD 4 M 9 Inframe del/ins c.1903_04delinsCCA/Val635delinsAlaMet NBF + + - P02* ccALD 5 M 9 Missense c.1979G.A/Arg660Gln - - 2.409 P03 ccALD 3 M IVS84 Frameshift g.1866-10G.A/Arg622fs Walker B3 - - P04 ccALD 4.5 M 1 Missense c.796G.A/Gly266Arg TMD + ++ 2.539 P05 ccALD 6 M 9 Frameshift c.1939_40insGG/Ala646fs NBF n.d - P06 ccALD 7 M 2 Missense c.904G.A/Glu302Lys TMD + + 2.194 P07 ccALD 8 M 3 Missense c.1202G.A/Arg401Gln - + ++ 2.396 P08* ccALD 8 M 10 Inframe del/ins c.1993_95delinsGAG/Lys665delinsGlu - + + - P09 AdolCALD 11 M 1 Missense c.796G.A/Gly266Arg TMD + ++ 2.539 P10 AdolCALD 11 M 8 Missense c.1816T.C/Ser606Pro C sequence - 2.499 P11 AdolCALD 15 M IVS8 Frameshift g.1866-10G.A/Arg622fs Walker B + - P12 ACALD 42 M 8 Missense c.1825G.A/Glu609Lys C sequence3 - 2.075 P13* ACALD 46 M 7 Missense c.1673T.C/Ile558Thr NBF3 + ++ 1.211 P14 AMN 26 M 9 Frameshift c.1939_40insGG/Ala646fs - - - P15 AMN 35 M 1 Missense c.796G.A/Gly266Arg TMD2 + ++ 2.539 P16 Asymptomatic 18 F 7 Missense c.1771C.T/Arg591Trp NBF + ++ 2.818 P17 Asymptomatic 26 F 1 Frameshift c.110_17del8/Val36fs - + + - *Novel Mutations, 1 ccALD-Childhood Adrenoleukodystrophy, AMN-Adrenomyeloneuropathy, ACALD-Adult Cerebral Adrenoleukodystrophy, AdolCALD- Adolescent cerebral Adrenoleukodystrophy.
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ABCD1 p.Ser606Pro 21966424:156:726
status: NEW168 Lane 1 and 10 (Control by symbol ''C``), Lane 2 (P01, ccALD, V635delins A & M), Lane 3 (P02, ccALD, R660Q), Lane 4 (P03, ccALD, R622fs), Lane 5 (P04, ccALD, G266R), Lane 6 (P06, ccALD, E302K), Lane 7 (P07, ccALD, R401Q), Lane 8 (P08, ccALD, K665delinsE), Lane 9 (P09, AdolCALD, G266R), Lane 11 (P10, AdolCALD, S606P), Lane 12 (P11, AdolCALD, R622fs), Lane 13 (P12, ACALD, E609K), Lane 14 (P13, ACALD, I558T), Lane 15 (P14, AMN, A646fs), Lane 16 (P15, AMN, G266R), Lane 17 (P16, asymptomatic female, R591W) and Lane 18 (P17, asymptomatic female,).
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ABCD1 p.Ser606Pro 21966424:168:310
status: NEW198 Supporting Information Supporting Information S1 Frequently occurring mutations in (a) intervening sequence 8 (g.1866-10G.A/Arg622fs, shown as C.T in antisense strand) in P03 and P11, (b) exon 1 (c.796G.A/ Gly266Arg, shown as C.T in antisense strand) in P04, P09 and P15, (c) exon 9 (c.1939_40insGG/Ala646fs, shown as CC in antisense strand) in P05 and P14, (d) exon 2 (c.904G.A/ Glu302Lys) in P06, (e) exon 3 (c.1202G.A/Arg401Gln) in P07, (f) exon 8 (c.1816T.C/Ser606Pro) in P10, (g) exon 8 (c.1825G.A/ Glu609Lys) in P12, (h) exon 7 (c.1771C.T/Arg591Trp) in P16 and (i) exon 1 (c.110_17del8/Val36fs) in P17.
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ABCD1 p.Ser606Pro 21966424:198:462
status: NEW102 Of the 10 non-recurrent mutations in our study group i.e a frameshift mutation c.110_117del8 (Val36fs) in exon 1 was identified in an asymptomatic heterozygous female and 5 missense mutations, c.904G.A (Glu302Lys) in exon 2, c.1202G.A (Arg401Gln) in exon 3, c.1771C.T (Arg591Trp) in exon 7, c.1816T.C (Ser606Pro) and c.1825G.A (Glu609Lys) in exon 8 were present in 5 different patients.
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ABCD1 p.Ser606Pro 21966424:102:302
status: NEW155 Patients Phenotype1 Age(Year) Sex Exon/IVS Mutation Type Mutations Protein Localization ALDP PSIC Score5 P01* ccALD 4 M 9 Inframe del/ins c.1903_04delinsCCA/Val635delinsAlaMet NBF + + - P02* ccALD 5 M 9 Missense c.1979G.A/Arg660Gln - - 2.409 P03 ccALD 3 M IVS84 Frameshift g.1866-10G.A/Arg622fs Walker B3 - - P04 ccALD 4.5 M 1 Missense c.796G.A/Gly266Arg TMD + ++ 2.539 P05 ccALD 6 M 9 Frameshift c.1939_40insGG/Ala646fs NBF n.d - P06 ccALD 7 M 2 Missense c.904G.A/Glu302Lys TMD + + 2.194 P07 ccALD 8 M 3 Missense c.1202G.A/Arg401Gln - + ++ 2.396 P08* ccALD 8 M 10 Inframe del/ins c.1993_95delinsGAG/Lys665delinsGlu - + + - P09 AdolCALD 11 M 1 Missense c.796G.A/Gly266Arg TMD + ++ 2.539 P10 AdolCALD 11 M 8 Missense c.1816T.C/Ser606Pro C sequence - 2.499 P11 AdolCALD 15 M IVS8 Frameshift g.1866-10G.A/Arg622fs Walker B + - P12 ACALD 42 M 8 Missense c.1825G.A/Glu609Lys C sequence3 - 2.075 P13* ACALD 46 M 7 Missense c.1673T.C/Ile558Thr NBF3 + ++ 1.211 P14 AMN 26 M 9 Frameshift c.1939_40insGG/Ala646fs - - - P15 AMN 35 M 1 Missense c.796G.A/Gly266Arg TMD2 + ++ 2.539 P16 Asymptomatic 18 F 7 Missense c.1771C.T/Arg591Trp NBF + ++ 2.818 P17 Asymptomatic 26 F 1 Frameshift c.110_17del8/Val36fs - + + - *Novel Mutations, 1 ccALD-Childhood Adrenoleukodystrophy, AMN-Adrenomyeloneuropathy, ACALD-Adult Cerebral Adrenoleukodystrophy, AdolCALD- Adolescent cerebral Adrenoleukodystrophy.
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ABCD1 p.Ser606Pro 21966424:155:726
status: NEW167 Lane 1 and 10 (Control by symbol ''C``), Lane 2 (P01, ccALD, V635delins A & M), Lane 3 (P02, ccALD, R660Q), Lane 4 (P03, ccALD, R622fs), Lane 5 (P04, ccALD, G266R), Lane 6 (P06, ccALD, E302K), Lane 7 (P07, ccALD, R401Q), Lane 8 (P08, ccALD, K665delinsE), Lane 9 (P09, AdolCALD, G266R), Lane 11 (P10, AdolCALD, S606P), Lane 12 (P11, AdolCALD, R622fs), Lane 13 (P12, ACALD, E609K), Lane 14 (P13, ACALD, I558T), Lane 15 (P14, AMN, A646fs), Lane 16 (P15, AMN, G266R), Lane 17 (P16, asymptomatic female, R591W) and Lane 18 (P17, asymptomatic female,).
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ABCD1 p.Ser606Pro 21966424:167:310
status: NEW197 Supporting Information Supporting Information S1 Frequently occurring mutations in (a) intervening sequence 8 (g.1866-10G.A/Arg622fs, shown as C.T in antisense strand) in P03 and P11, (b) exon 1 (c.796G.A/ Gly266Arg, shown as C.T in antisense strand) in P04, P09 and P15, (c) exon 9 (c.1939_40insGG/Ala646fs, shown as CC in antisense strand) in P05 and P14, (d) exon 2 (c.904G.A/ Glu302Lys) in P06, (e) exon 3 (c.1202G.A/Arg401Gln) in P07, (f) exon 8 (c.1816T.C/Ser606Pro) in P10, (g) exon 8 (c.1825G.A/ Glu609Lys) in P12, (h) exon 7 (c.1771C.T/Arg591Trp) in P16 and (i) exon 1 (c.110_17del8/Val36fs) in P17.
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ABCD1 p.Ser606Pro 21966424:197:462
status: NEW[hide] [Adrenoleukodystrophy: structure and function of A... Yakugaku Zasshi. 2007 Jan;127(1):163-72. Takahashi N, Morita M, Imanaka T
[Adrenoleukodystrophy: structure and function of ALDP, and intracellular behavior of mutant ALDP with naturally occurring missense mutations].
Yakugaku Zasshi. 2007 Jan;127(1):163-72., [PMID:17202797]
Abstract [show]
Adrenoleukodystrophy (ALD) is an inherited disorder characterized by progressive demyelination of the central nervous system and adrenal dysfunction. The biochemical characterization is based on the accumulation of pathgnomonic amounts of saturated very long-chain fatty acid (VLCFA; C>22) in all tissues, including the brain white matter, adrenal glands, and skin fibroblasts, of the patients. The accumulation of VLCFA in ALD is linked to a mutation in the ALD (ABCD1) gene, an ABC subfamily D member. The ALD gene product, so-called ALDP (ABCD1), is thought to be involved in the transport of VLCFA or VLCFA-CoA into the peroxisomes. ALDP is a half-sized peroxisomal ABC protein and it has 745 amino acids in humans. ALDP is thought to be synthesized on free polysomes, posttranslationally transported to peroxisomes, and inserted into the membranes. During this process, ALDP interacts with Pex19p, a chaperone-like protein for intracellular trafficking of peroxisomal membrane protein (PMP), the complex targets Pex3p on the peroxisomal membranes, and ALDP is inserted into the membranes. After integration into the membranes, ALDP is thought to form mainly homodimers. Here, we chose nine arbitrary mutations of human ALDP with naturally occurring missense mutations and examined the intracellular behavior of their ALDPs. We found that mutant ALDP (S606L, R617H, and H667D) was degraded together with wild-type ALDP by proteasomes. These results suggest that the complex of mutant and wild-type ALDP is recognized as misfolded proteins and degraded by the protein quality control system associated with proteasomes. Further, we found fragmentation of mutant ALDP (R104C) on peroxisomes and it was not inhibited by proteasomes inhibitors, suggesting that an additional protease(s) is also involved in the quality control of mutant ALDP. In addition, mutation of ALDP (Y174C) suggests that a loop between transmembrane domains 2 and 3 is important for the targeting of ALDP to peroxisomes.
Comments [show]
None has been submitted yet.
No. Sentence Comment
19 S606P and S606L are in ABC signature motif.
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ABCD1 p.Ser606Pro 17202797:19:0
status: NEW28 ミスセンス変異を持つ ALDP の細胞内動態 ―一過性発3fe;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.
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ABCD1 p.Ser606Pro 17202797:28:179
status: NEWX
ABCD1 p.Ser606Pro 17202797:28:1381
status: NEWX
ABCD1 p.Ser606Pro 17202797:28:5742
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.
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ABCD1 p.Ser606Pro 17202797:29:180
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.
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ABCD1 p.Ser606Pro 17202797:36:934
status: NEWX
ABCD1 p.Ser606Pro 17202797:36:1532
status: NEWX
ABCD1 p.Ser606Pro 17202797:36:2516
status: NEWX
ABCD1 p.Ser606Pro 17202797:36:3869
status: NEWX
ABCD1 p.Ser606Pro 17202797:36:5036
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.
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ABCD1 p.Ser606Pro 17202797:43:1473
status: NEWX
ABCD1 p.Ser606Pro 17202797:43:4720
status: NEW49 変異型 ALDP の分9e3;過程の解析 新生タンパク質が正しい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.
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ABCD1 p.Ser606Pro 17202797:49:50
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).
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ABCD1 p.Ser606Pro 17202797:52:50
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.
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ABCD1 p.Ser606Pro 17202797:53:701
status: NEWX
ABCD1 p.Ser606Pro 17202797:53:2620
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.Ser606Pro 17202797:27:1221
status: NEWX
ABCD1 p.Ser606Pro 17202797:27:5079
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.Ser606Pro 17202797:34:830
status: NEWX
ABCD1 p.Ser606Pro 17202797:34:1361
status: NEWX
ABCD1 p.Ser606Pro 17202797:34:2235
status: NEWX
ABCD1 p.Ser606Pro 17202797:34:3428
status: NEWX
ABCD1 p.Ser606Pro 17202797:34:3644
status: NEWX
ABCD1 p.Ser606Pro 17202797:34:4461
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.
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ABCD1 p.Ser606Pro 17202797:41:1307
status: NEWX
ABCD1 p.Ser606Pro 17202797:41:4175
status: NEW[hide] Mutations, clinical findings and survival estimate... PLoS One. 2012;7(3):e34195. doi: 10.1371/journal.pone.0034195. Epub 2012 Mar 29. Pereira Fdos S, Matte U, Habekost CT, de Castilhos RM, El Husny AS, Lourenco CM, Vianna-Morgante AM, Giuliani L, Galera MF, Honjo R, Kim CA, Politei J, Vargas CR, Jardim LB
Mutations, clinical findings and survival estimates in South American patients with X-linked adrenoleukodystrophy.
PLoS One. 2012;7(3):e34195. doi: 10.1371/journal.pone.0034195. Epub 2012 Mar 29., [PMID:22479560]
Abstract [show]
In this study, we analyzed the ABCD1 gene in X-linked adrenoleukodystrophy (X-ALD) patients and relatives from 38 unrelated families from South America, as well as phenotypic proportions, survival estimates, and the potential effect of geographical origin in clinical characteristics. METHODS: X- ALD patients from Brazil, Argentina and Uruguay were invited to participate in molecular studies to determine their genetic status, characterize the mutations and improve the genetic counseling of their families. All samples were screened by SSCP analysis of PCR fragments, followed by automated DNA sequencing to establish the specific mutation in each family. Age at onset and at death, male phenotypes, genetic status of women, and the effect of family and of latitude of origin were also studied. RESULTS: We identified thirty-six different mutations (twelve novel). This population had an important allelic heterogeneity, as only p.Arg518Gln was repeatedly found (three families). Four cases carried de novo mutations. Intra-familiar phenotype variability was observed in all families. Out of 87 affected males identified, 65% had the cerebral phenotype (CALD). The mean (95% CI) ages at onset and at death of the CALD were 10.9 (9.1-12.7) and 24.7 (19.8-29.6) years. No association was found between phenotypic manifestations and latitude of origin. One index-case was a girl with CALD who carried an ABCD1 mutation, and had completely skewed X inactivation. CONCLUSIONS: This study extends the spectrum of mutations in X-ALD, confirms the high rates of de novo mutations and the absence of common mutations, and suggests a possible high frequency of cerebral forms in our population.
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No. Sentence Comment
26 NBF Inherited Northern Brazil 31/Male CALD p.Arg518Gln (Imamura A et al., 1997) E6 Missense c.1553G.A CGG.CAG NBF de novo Southern Brazil 32/Male CALD p.Arg401Trp (Takano H et al., 1999) E3 Missense c.1201C.T CGG.UGG - ND Southern Brazil 33/Male CALD p.Thr632Pro (http://www.x-ald.nl) E9 Missense c.1894A.C ACC.CCC NBF de novo Southern Brazil 36/Male CALD p.Arg518Gln (Imamura A et al., 1997) E6 Missense c.1553G.A CGG.CAG NBF Inherited Northern Brazil 37/Male CALD p.Ser358X (Coll MJ et al., 2005) E2 Stop codon c.1073C.G UCA.UGA TMD Inherited Southern Brazil 38/Male CALD p.Ile481Phe # E5 Missense c.1441A.T AUC.UUC NBF Inherited Northern Brazil 39/Male AMN p.Arg389Gly (Krasemann EW et al., 1996) E3 Missense c.1165C.G CGC.GGC - ND Argentina 40/Male AMN p.Gln472fsX83 (Barcelo &#b4; A et al., 1994) E5 Frameshift+stop codon c.1415_1416delAG Truncated - Inherited Uruguay 41/Male CALD p.Ala95fsX11 # E1B Frameshift+stop codon c.283_284ins9 Elonged TMD Inherited Southern Brazil 44/Male CALD p.Ser606Pro (Feigenbaum V et al. 1996) E8 Missense c.1816T.C UCG.CCG NBF Inherited Northern Brazil 45/Male CALD p.Gln55X # E1A Stop codon c.163C.T Truncated - Inherited Northern Brazil 46/Male CALD p.Glu199Lys (http://www.x-ald.nl) E1C Missense c.595G.A GAG.AAG TMD ND Northern Brazil common central demyelinative disease.
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ABCD1 p.Ser606Pro 22479560:26:995
status: NEW