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. 1995 Aug;57(2):292–301.

Altered expression of ALDP in X-linked adrenoleukodystrophy.

P A Watkins 1, S J Gould 1, M A Smith 1, L T Braiterman 1, H M Wei 1, F Kok 1, A B Moser 1, H W Moser 1, K D Smith 1
PMCID: PMC1801558  PMID: 7668254

Abstract

X-linked adrenoleukodystrophy (ALD) is a neurodegenerative disorder with variable phenotypic expression that is characterized by elevated plasma and tissue levels of very long-chain fatty acids. However, the product of the gene defective in ALD (ALDP) is a membrane transporter of the ATP-binding cassette family of proteins and is not related to enzymes known to activate or oxidize fatty acids. We generated an antibody that specifically recognizes the C-terminal 18 amino acids of ALDP and can detect ALDP by indirect immunofluorescence. To better understand the mechanism by which mutations in ALDP lead to disease, we used this antibody to examine the subcellular distribution and relative abundance of ALDP in skin fibroblasts from normal individuals and ALD patients. Punctate immunoreactive material typical of fibroblast peroxisomes was observed in cells from seven normal controls and eight non-ALD patients. Of 35 ALD patients tested, 17 had the childhood-onset cerebral form of the disease, 13 had the milder adult phenotype adrenomyeloneuropathy, 3 had adrenal insufficiency only, and 2 were affected fetuses. More than two-thirds (69%) of all patients studied showed no punctate immunoreactive material. There was no correlation between the immunofluorescence pattern and clinical phenotype. We determined the mutation in the ALD gene in 15 of these patients. Patients with either a deletion or frameshift mutation lacked ALDP immunoreactivity, as expected. Four of 11 patients with missense mutations were also immunonegative, indicating that these mutations affected the stability or localization of ALDP. In the seven immunopositive patients with missense mutations, correlation of the location and nature of the amino acid substitution may provide new insights into the function of this peroxisomal membrane protein. Furthermore, the study of female relatives of immunonegative ALD probands may aid in the assessment of heterozygote status.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berger J., Molzer B., Faé I., Bernheimer H. X-linked adrenoleukodystrophy (ALD): a novel mutation of the ALD gene in 6 members of a family presenting with 5 different phenotypes. Biochem Biophys Res Commun. 1994 Dec 30;205(3):1638–1643. doi: 10.1006/bbrc.1994.2855. [DOI] [PubMed] [Google Scholar]
  2. Cartier N., Sarde C. O., Douar A. M., Mosser J., Mandel J. L., Aubourg P. Abnormal messenger RNA expression and a missense mutation in patients with X-linked adrenoleukodystrophy. Hum Mol Genet. 1993 Nov;2(11):1949–1951. doi: 10.1093/hmg/2.11.1949. [DOI] [PubMed] [Google Scholar]
  3. Fanen P., Guidoux S., Sarde C. O., Mandel J. L., Goossens M., Aubourg P. Identification of mutations in the putative ATP-binding domain of the adrenoleukodystrophy gene. J Clin Invest. 1994 Aug;94(2):516–520. doi: 10.1172/JCI117363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fuchs S., Sarde C. O., Wedemann H., Schwinger E., Mandel J. L., Gal A. Missense mutations are frequent in the gene for X-chromosomal adrenoleukodystrophy (ALD). Hum Mol Genet. 1994 Oct;3(10):1903–1905. doi: 10.1093/hmg/3.10.1903. [DOI] [PubMed] [Google Scholar]
  5. Gould S. J., Krisans S., Keller G. A., Subramani S. Antibodies directed against the peroxisomal targeting signal of firefly luciferase recognize multiple mammalian peroxisomal proteins. J Cell Biol. 1990 Jan;110(1):27–34. doi: 10.1083/jcb.110.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Igarashi M., Schaumburg H. H., Powers J., Kishmoto Y., Kolodny E., Suzuki K. Fatty acid abnormality in adrenoleukodystrophy. J Neurochem. 1976 Apr;26(4):851–860. doi: 10.1111/j.1471-4159.1976.tb04462.x. [DOI] [PubMed] [Google Scholar]
  7. Janski A. M., Cornell N. W. Subcellular distribution of enzymes determined by rapid digitonin fractionation of isolated hepatocytes. Biochem J. 1980 Feb 15;186(2):423–429. doi: 10.1042/bj1860423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Keller G. A., Gould S., Deluca M., Subramani S. Firefly luciferase is targeted to peroxisomes in mammalian cells. Proc Natl Acad Sci U S A. 1987 May;84(10):3264–3268. doi: 10.1073/pnas.84.10.3264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kemp S., Ligtenberg M. J., van Geel B. M., Barth P. G., Wolterman R. A., Schoute F., Sarde C. O., Mandel J. L., van Oost B. A., Bolhuis P. A. Identification of a two base pair deletion in five unrelated families with adrenoleukodystrophy: a possible hot spot for mutations. Biochem Biophys Res Commun. 1994 Jul 29;202(2):647–653. doi: 10.1006/bbrc.1994.1979. [DOI] [PubMed] [Google Scholar]
  10. Köster W., Böhm B. Point mutations in two conserved glycine residues within the integral membrane protein FhuB affect iron(III) hydroxamate transport. Mol Gen Genet. 1992 Apr;232(3):399–407. doi: 10.1007/BF00266243. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lazo O., Contreras M., Hashmi M., Stanley W., Irazu C., Singh I. Peroxisomal lignoceroyl-CoA ligase deficiency in childhood adrenoleukodystrophy and adrenomyeloneuropathy. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7647–7651. doi: 10.1073/pnas.85.20.7647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ligtenberg M. J., Kemp S., Sarde C. O., van Geel B. M., Kleijer W. J., Barth P. G., Mandel J. L., van Oost B. A., Bolhuis P. A. Spectrum of mutations in the gene encoding the adrenoleukodystrophy protein. Am J Hum Genet. 1995 Jan;56(1):44–50. [PMC free article] [PubMed] [Google Scholar]
  14. Matsumoto T., Kondoh T., Masuzaki H., Harada N., Matsusaka T., Kinoshita E., Takeo G., Tsujihata M., Suzuki Y., Tsuji Y. A point mutation at ATP-binding region of the ALD gene in a family with X-linked adrenoleukodystrophy. Jpn J Hum Genet. 1994 Sep;39(3):345–351. doi: 10.1007/BF01874053. [DOI] [PubMed] [Google Scholar]
  15. McCombie W. R., Heiner C., Kelley J. M., Fitzgerald M. G., Gocayne J. D. Rapid and reliable fluorescent cycle sequencing of double-stranded templates. DNA Seq. 1992;2(5):289–296. doi: 10.3109/10425179209030961. [DOI] [PubMed] [Google Scholar]
  16. McGuinness M. C., Moser A. B., Poll-The B. T., Watkins P. A. Complementation analysis of patients with intact peroxisomes and impaired peroxisomal beta-oxidation. Biochem Med Metab Biol. 1993 Apr;49(2):228–242. doi: 10.1006/bmmb.1993.1025. [DOI] [PubMed] [Google Scholar]
  17. Moser H. W., Moser A. B., Frayer K. K., Chen W., Schulman J. D., O'Neill B. P., Kishimoto Y. Adrenoleukodystrophy: increased plasma content of saturated very long chain fatty acids. Neurology. 1981 Oct;31(10):1241–1249. doi: 10.1212/wnl.31.10.1241. [DOI] [PubMed] [Google Scholar]
  18. Moser H. W., Moser A. B., Kawamura N., Murphy J., Suzuki K., Schaumburg H., Kishimoto Y. Adrenoleukodystrophy: elevated C26 fatty acid in cultured skin fibroblasts. Ann Neurol. 1980 Jun;7(6):542–549. doi: 10.1002/ana.410070607. [DOI] [PubMed] [Google Scholar]
  19. Mosser J., Douar A. M., Sarde C. O., Kioschis P., Feil R., Moser H., Poustka A. M., Mandel J. L., Aubourg P. Putative X-linked adrenoleukodystrophy gene shares unexpected homology with ABC transporters. Nature. 1993 Feb 25;361(6414):726–730. doi: 10.1038/361726a0. [DOI] [PubMed] [Google Scholar]
  20. Mosser J., Lutz Y., Stoeckel M. E., Sarde C. O., Kretz C., Douar A. M., Lopez J., Aubourg P., Mandel J. L. The gene responsible for adrenoleukodystrophy encodes a peroxisomal membrane protein. Hum Mol Genet. 1994 Feb;3(2):265–271. doi: 10.1093/hmg/3.2.265. [DOI] [PubMed] [Google Scholar]
  21. Saurin W., Köster W., Dassa E. Bacterial binding protein-dependent permeases: characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins. Mol Microbiol. 1994 Jun;12(6):993–1004. doi: 10.1111/j.1365-2958.1994.tb01087.x. [DOI] [PubMed] [Google Scholar]
  22. Singh H., Derwas N., Poulos A. Very long chain fatty acid beta-oxidation by rat liver mitochondria and peroxisomes. Arch Biochem Biophys. 1987 Dec;259(2):382–390. doi: 10.1016/0003-9861(87)90504-2. [DOI] [PubMed] [Google Scholar]
  23. Singh H., Poulos A. Distinct long chain and very long chain fatty acyl CoA synthetases in rat liver peroxisomes and microsomes. Arch Biochem Biophys. 1988 Nov 1;266(2):486–495. doi: 10.1016/0003-9861(88)90281-0. [DOI] [PubMed] [Google Scholar]
  24. Singh I., Moser A. E., Goldfischer S., Moser H. W. Lignoceric acid is oxidized in the peroxisome: implications for the Zellweger cerebro-hepato-renal syndrome and adrenoleukodystrophy. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4203–4207. doi: 10.1073/pnas.81.13.4203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith L. M., Sanders J. Z., Kaiser R. J., Hughes P., Dodd C., Connell C. R., Heiner C., Kent S. B., Hood L. E. Fluorescence detection in automated DNA sequence analysis. Nature. 1986 Jun 12;321(6071):674–679. doi: 10.1038/321674a0. [DOI] [PubMed] [Google Scholar]
  26. Swinkels B. W., Gould S. J., Bodnar A. G., Rachubinski R. A., Subramani S. A novel, cleavable peroxisomal targeting signal at the amino-terminus of the rat 3-ketoacyl-CoA thiolase. EMBO J. 1991 Nov;10(11):3255–3262. doi: 10.1002/j.1460-2075.1991.tb04889.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Uchiyama A., Suzuki Y., Song X. Q., Fukao T., Imamura A., Tomatsu S., Shimozawa N., Kondo N., Orii T. Identification of a nonsense mutation in ALD protein cDNA from a patient with adrenoleukodystrophy. Biochem Biophys Res Commun. 1994 Jan 28;198(2):632–636. doi: 10.1006/bbrc.1994.1092. [DOI] [PubMed] [Google Scholar]
  28. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wanders R. J., van Roermund C. W., van Wijland M. J., Schutgens R. B., van den Bosch H., Schram A. W., Tager J. M. Direct demonstration that the deficient oxidation of very long chain fatty acids in X-linked adrenoleukodystrophy is due to an impaired ability of peroxisomes to activate very long chain fatty acids. Biochem Biophys Res Commun. 1988 Jun 16;153(2):618–624. doi: 10.1016/s0006-291x(88)81140-9. [DOI] [PubMed] [Google Scholar]

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