Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Cloning of the human aspartoacylase cDNA and a common missense mutation in Canavan disease

Nature Genetics volume 5, pages 118–123 (1993)Cite this article

Abstract

Canavan disease, an autosomal recessive leukodystrophy, is caused by deficiency of aspartoacylase and accumulation of N–acetylaspartic acid in brain. We have cloned the human aspartoacylase (ASP) cDNA spanning 1,435 basepairs, and show that the isolated cDNA expresses aspartoacylase activity in bacteria. Furthermore, an A to C base change, at nucleotide 854, has been found in 85% of the 34 Canavan alleles tested so far. This base change results in a missense Glu285Ala mutation that is predicted to be part of the catalytic domain of aspartoacylase. The data suggest that the catalytic centre of aspartoacylase involves a triad of Ser, His and Glu residues. Our findings have implications for diagnosis and screening of Canavan disease.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Globus, J.H. & Strauss, I. Progressive degenerative subcortical encephalopathy (Schilder's disease). Arch. Neurol. Psychiat. 20, 1190–1228 (1928).

    Article  Google Scholar 

  2. Canavan, M.M. Schilder's encephalitis periaxialis diffusa. Arch. Neurol. Psychiat 25, 299–308 (1931).

    Article  Google Scholar 

  3. van Bogaert, L. & Bertrand, I. Sur une idiotie familiale avec degerescence sponglieuse de neuraxe (note preliminaire). Acta. Neurol. Belg. 49, 572–587 (1949).

    Google Scholar 

  4. Adachi, M., Torii, J., Schneck, L. & Volk, B.W. Electron microscopic and enzyme histochemical studies of the cerebellum in spongy degeneration (van Bogaert and Bertrand type). Acta. Neuropath. 20, 22–31 (1972).

    Article  CAS  Google Scholar 

  5. Adornato, B.T., O'Brien, J.S., Lamport, P.W., Roe, T.F. & Neustein, H.B. Cerebral spongy degeneration of infancy: a biochemical and ultrastructural study of affected twins. Neurology 22, 202–210 (1972).

    Article  CAS  Google Scholar 

  6. Matalon, R., Michals, K., Sebasta, D., Deanching, M., Gashkoff, P. & Casanova, J. Aspartoacylase deficiency and N-acetylaspartic aciduria in patients with Canavan Disease. Am. J. med. Genet. 29, 463–471 (1988).

    Article  CAS  Google Scholar 

  7. Bimbaum, S.M. Amino acid acylases I and II from hog kidney. Meth. Enzymol. 2, 115–119 (1955).

    Article  CAS  Google Scholar 

  8. Bimbaum, S.M., Levinton, L., Kingsley, R.B. & Greenstein, J.P. Specificity of amino acid acylases. J. biol. Chem. 194, 455–462 (1952).

    Google Scholar 

  9. Matalon, R. et al. Aspartoacylase deficiency: the enzyme defect in Canavan Disease. J. inher. met. Dis. 12, 329–331 (1989).

    Article  Google Scholar 

  10. Matalon, R., Kaul, R. & Michals, K. Canavan disease: biochemical and molecular studies. J. inher. metab. Dis. (in the press).

  11. Grodd, W., Krägeloh-Mann, I., Petersen, D., Trefz, F.K. & Harzer, K. In vivo asessment of N-acetylaspartate in brain in spongy degeneration (Canavan disease) by proton spectroscopy. Lancet 336, 437–438 (1990).

    Article  CAS  Google Scholar 

  12. Matalon, R., Kaul, R. & Michals, K. . In The Molecular Biology and Genetic Basis of Neurological Disease (eds Rosenberg, R.N. et al.) 541–546 (Butterworth-Heineman, Boston, 1993).

    Google Scholar 

  13. Ozand, P.T., Gascon, G. & Dhalla, M. Aspartoacylase deficiency and canavan disease in Saudi Arabia. Am. J. med. Genet. 35, 266–268 (1990).

    Article  CAS  Google Scholar 

  14. Michelakakis, H., Giouroukos, S., Divry, P., Katsarou, E., Rolland, M.O. & Skardoutsow, A., Canavan Disease: findings in four new cases. J. inher. metab. Dis. 14, 267–268 (1991).

    Article  CAS  Google Scholar 

  15. Matalon, R., Michals, K., Gashkoff, P. & Kaul, R. Prenatal diagnosis of Canavan disease. J. inher. metab. Dis. 15, 392–394 (1992).

    Article  CAS  Google Scholar 

  16. Kaul, R., Casanova, J., Johnson, A., Tang, P. & Matalon, R. Purification, characterization and localization of aspartoacylase from bovine brain. J. Neurochem. 56, 129–135 (1991).

    Article  CAS  Google Scholar 

  17. Proudfoot, N.J. & Brownlee, G.G. 3′ Non-coding region sequences in eukaryotic messenger RNA. Nature 263, 211–214 (1976).

    Article  CAS  Google Scholar 

  18. Birnstiel, M.L., Busslinger, M. & Strub, K. Transcription termination and 3′ processing: the end is in site! Cell 41, 349–359 (1985).

    Article  CAS  Google Scholar 

  19. Cygler, M. et al. Relationship between sequence conservation and three-dimentional structure in a large family of esterases, lipases, and related proteins. Protein Sci. 2, 366–382 (1993).

    Article  CAS  Google Scholar 

  20. Lemeur, M.A., Galliot, B. & Geriinger, P. Termination of the ovalbumin gene transcription. EMBO J. 3, 2779–2786 (1984).

    Article  CAS  Google Scholar 

  21. Miller, Y.R., Drabkin, H., Jones, C. & Fisher, J.H. Human aminoacylase-1: cloning, regional assignment to distal chromosome 3p21.1, and identification of a cross-hybridizing sequence on chromosome 18. Genomics 8, 149–154 (1990).

    Article  CAS  Google Scholar 

  22. Goldstein, F.B. Biosynthesis of N-acetyl-L-aspartic acid. J. biol. Chem. 234, 2702–2706 (1959).

    CAS  Google Scholar 

  23. Goldstein, F.B. The enzymatic synthesis of N-acetyl-L-aspartic acid by subcellular preparations of rat brain. J. biol. Chem. 244, 4257–4260 (1969).

    CAS  PubMed  Google Scholar 

  24. Moffett, J.R., Namboodiri, M.A., Cangro, C.B. & Neale, J.H. Immunohistochemical localization of N-acetylaspartate in rat brain. NeuroReport 2, 131–134 (1991).

    Article  CAS  Google Scholar 

  25. Urenjak, J., Williams, S.R., Gadian, D.G. & Noble, M. Specific expression of N-acetylaspartate in neurons, oligodendrocyte-type-2 astrocyte progenitors, and immature oligodendrocytes in vitro. J. Neurochem. 59, 55–61 (1992).

    Article  CAS  Google Scholar 

  26. Moffett, J.R., Namboodiri, M.A. & Neale, J.H. Enhanced carbodiimide fixation for immunohistochemistry: application to the comparative distributions of N-acetylaspartylglutamate and N-acetylaspartate immunoreactivities in rat brain. J. Histochem. Cytochem. 31, 559–570 (1993).

    Article  Google Scholar 

  27. Tallan, H.H., Moore, S. & Stein, W.H. N-Acetyl-L-aspartic acid in brain. J. biol. Chem. 219, 257–264 (1956).

    CAS  PubMed  Google Scholar 

  28. Jacobson, K.B. Studies on the role of N-acetylaspartic acid on mammalian brain. J. gen. Physiol. 43, 323–333 (1957).

    Article  Google Scholar 

  29. Mclntosh, J.M. & Cooper, J.R. Studies on the function of N-acetylaspartic acid in the brain. J. Neurochem. 12, 825–835 (1965).

    Article  Google Scholar 

  30. Shigematsu, H. et al. Purification and characterization of the heat stable factors essential for conversion of lignoceric acid to cerebronic acid and glutamic acid: identification of N-acetyl-L-aspartic acid. J. Neurochem. 40, 814–620 (1983).

    Article  CAS  Google Scholar 

  31. Cangro, C.B., Namboodiri, M.A.A., Sklae, L.A., Corigliano-Murphy, A. & Neale, J.H. Immunocytochemistry and biosynthesis of N-acetylaspartylglutamate in spinal sensory ganglia. J. Neurochem. 49, 1579–1588 (1987).

    Article  CAS  Google Scholar 

  32. Ory-Lavollee, L., Blakely, R.D. & Coyle, J.T. Neurochemical and immunocytochemical studies on the distribution of N-acetylaspartylglutamate and N-acetyl-aspartate in rat spinal cord and some peripheral nervous tissues. J. Neurochem. 48, 895–899 (1987).

    Article  CAS  Google Scholar 

  33. Birken, D.L. & Oldendorf, W.H. N-Acetyl-L-aspartic acid: A literature review of a compount prominent in 1H-NMR spectroscopic studies on brain. Neurosci. Behavioral Rev. 13, 23–31 (1989).

    Article  CAS  Google Scholar 

  34. Grood, W., Krägeloh-Mann, I., Klose, U. & Sauter, R. Metabolic and destructive brain disorders in children: Finding with localized proton MR spectroscopy. Radiology 181, 173–181 (1991).

    Article  Google Scholar 

  35. Dunlop, D.S., McHale, D.M. & Lajtha, A. Decreased brain N-acetylaspartate in Huntingdon's disease. Brain Res. 580, 44–48 (1992).

    Article  CAS  Google Scholar 

  36. Meyerhoff, D.J. et al. Reduced brain N- acetylaspartate suggests neuronal loss in cognitively impaired human immunodeficiency virus-seropositive individuals: In vivo 1H magnetic resonance spectroscopic imaging. Neurology 43, 509–515 (1993).

    Article  CAS  Google Scholar 

  37. Gideon, P. et al. Early time course of N-acetylspartate, creatine and phosphocreatine, and compounds containing choline in the brain after acute stroke-a proton magnetic resonance spectroscopy study. Stroke 23, 1566–1572 (1992).

    Article  CAS  Google Scholar 

  38. Marcucci, F., Colombo, L., De Ponte, G. & Mussini, E. Decrease in N-acetyl-L-aspartic acid in brain of myodystrophic mice. J. Neurochem. 43, 1484–1486 (1984).

    Article  CAS  Google Scholar 

  39. Bell, J.D. et al. In vivo detection of metabolic changes in a mouse model of scrapie using nuclear magnetic resonance spectroscopy. J. gen. Virology 72, 2419–2423 (1991).

    Article  CAS  Google Scholar 

  40. Kaul, R., Murthy, S.N.P., Reddy, A.G., Steck, T.L. & Kohler, H. Amino acid sequence of the N-terminal 201 residues of human erythrocyte membrane band 3. J. biol. Chem. 258, 7981–7990 (1983).

    CAS  PubMed  Google Scholar 

  41. Saiki, R.K. et al. Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230, 1350–1354 (1985).

    Article  CAS  Google Scholar 

  42. Orita, M., Suzuki, Y., Sekiya, T. & Hayashi, K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5, 874–879 (1989).

    Article  CAS  Google Scholar 

  43. Feinberg, A.P. & Vogelstein, B. Technique for radio labelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137, 266–267 (1984).

    Article  CAS  Google Scholar 

  44. Kaul, R., Hidebrand, B., Roberts, S. & Jagadeeswaran, P. Isolation and characeterization of human blood coagulation factor X cDNA. Gene 41, 311–314 (1986).

    Article  CAS  Google Scholar 

  45. Sambrook, J., Fritsch, E.F. & Maniatis, T. Molecular Cloning: A laboratory manual 2nd edn (Cold Spring Harbour Laboratory, New York, 1989).

    Google Scholar 

  46. Sanger, F.S., Nicklen, I. & Coulson, A.R. DNA sequencing with chain terminating inhibitors. Proc. natn. Acad. Sci. U.S.A. 74, 5463–5467 (1977).

    Article  CAS  Google Scholar 

  47. Chomczynski, P. & Sacchi, N. Single step method of RNA isolation by acid guanidiniumthiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156 (1987).

    Article  CAS  Google Scholar 

  48. Lowry, O.H., Rosenbroug, N.J., Farr, A.L. & Randall, R.J. Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951).

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research Institute, Miami Children's Hospital, Miami, Florida, 33155, USA

    Rajinder Kaul, Guang Ping Gao, Kuppareddi Balamurugan & Reuben Matalon

Authors
  1. Rajinder Kaul
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guang Ping Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kuppareddi Balamurugan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Reuben Matalon
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kaul, R., Ping Gao, G., Balamurugan, K. et al. Cloning of the human aspartoacylase cDNA and a common missense mutation in Canavan disease. Nat Genet 5, 118–123 (1993). https://doi.org/10.1038/ng1093-118

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1093-118

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing