Impaired very long-chain acyl-CoA ß-oxidation in human X-linked adrenoleukodystrophy fibroblasts is a direct consequence of ABCD1 transporter dysfunction.
J Biol Chem
; 288(26): 19269-79, 2013 Jun 28.
Article
em En
| MEDLINE
| ID: mdl-23671276
ABSTRACT
X-linked adrenoleukodystrophy (X-ALD), an inherited peroxisomal disorder, is caused by mutations in the ABCD1 gene encoding the peroxisomal ATP-binding cassette (ABC) transporter ABCD1 (adrenoleukodystrophy protein, ALDP). Biochemically, X-ALD is characterized by an accumulation of very long-chain fatty acids and partially impaired peroxisomal ß-oxidation. In this study, we used primary human fibroblasts from X-ALD and Zellweger syndrome patients to investigate the peroxisomal ß-oxidation defect. Our results show that the degradation of C260-CoA esters is as severely impaired as degradation of unesterified very long-chain fatty acids in X-ALD and is abolished in Zellweger syndrome. Interestingly, the ß-oxidation rates for both C260-CoA and C220-CoA were similarly affected, although C220 does not accumulate in patient fibroblasts. Furthermore, we show that the ß-oxidation defect in X-ALD is directly caused by ABCD1 dysfunction as blocking ABCD1 function with a specific antibody reduced ß-oxidation to levels observed in X-ALD fibroblasts. By quantification of mRNA and protein levels of the peroxisomal ABC transporters and by blocking with specific antibodies, we found that residual ß-oxidation activity toward C260-CoA in X-ALD fibroblasts is mediated by ABCD3, although the efficacy of ABCD3 appeared to be much lower than that of ABCD1. Finally, using isolated peroxisomes, we show that ß-oxidation of C260-CoA is independent of additional CoA but requires a cytosolic factor of >10-kDa molecular mass that is resistant to N-ethylmaleimide and heat inactivation. In conclusion, our findings in human cells suggest that, in contrast to yeast cells, very long-chain acyl-CoA esters are transported into peroxisomes by ABCD1 independently of additional synthetase activity.
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Base de dados:
MEDLINE
Assunto principal:
Acil Coenzima A
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Transportadores de Cassetes de Ligação de ATP
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Adrenoleucodistrofia
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Fibroblastos
Limite:
Humans
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Male
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article