Human Genetics

, Volume 100, Issue 3–4, pp 450–456

Adult onset globoid cell leukodystrophy (Krabbe disease): analysis of galactosylceramidase cDNA from four Japanese patients

  • H. Furuya
  • Yoh-ji Kukita
  • Sukehisa Nagano
  • Yasuyoshi Sakai
  • Yoriaki Yamashita
  • Hidenao Fukuyama
  • Yuichiro Inatomi
  • Yutaka Saito
  • Ryoko Koike
  • Shoji Tsuji
  • Yasuyuki Fukumaki
  • Kenshi Hayashi
  • Takuro Kobayashi
Original investigation

Abstract

We examined galactosylceramidase (GALC) cDNA in four Japanese patients with adult onset globoid cell leukodystrophy (Krabbe disease; AO-GLD) by polymerase chain reaction/single-strand conformation polymorphism (PCR-SSCP) analysis, subsequent sequence determination, and restriction enzyme digestion of PCR products. Initial symptoms were the onset of slowly progressive spastic paraplegia from the middle of the second decade, and all patients had diminished GALC activity in their leukocytes. We identified three missense mutations (I66M, G270D, L618S) and one exon-6 skipping (535– 573del). Two of the patients had only the I66M mutant mRNA, and one only the G270D mutant mRNA. The fourth patient carried a compound heterozygous mutation of 535–573del and L618S. To determine the enzymatic activities produced by these mutations, we constructed mutated GALC cDNAs and expressed them in COS-1 cells. Three mutations, viz., G270D, L618S, and exon-6 skipping (535–573del), produced diminished GALC activity as expected. The I66M mutation in the wild-type GALC cDNA(I289) had normal activity, but when this mutation and the V289 polymorphism were introduced into the same allele, it had decreased activity. Thus, the combination of a unique mutation and polymorphism causes conformational change in the GALC enzyme, resulting in low enzymatic activity. AO-GLD mutations, including those found here, are located in the N-terminus (I66M, G270D, 535–573del) or C-terminus (L618S) of the GALC enzyme, whereas the reported mutations in the infantile form (IF-GLD) are in the central domain. This difference in mutation sites may affect the clinical features of GLD.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • H. Furuya
    • 1
  • Yoh-ji Kukita
    • 2
  • Sukehisa Nagano
    • 1
  • Yasuyoshi Sakai
    • 3
  • Yoriaki Yamashita
    • 4
  • Hidenao Fukuyama
    • 5
  • Yuichiro Inatomi
    • 6
  • Yutaka Saito
    • 7
  • Ryoko Koike
    • 7
  • Shoji Tsuji
    • 7
  • Yasuyuki Fukumaki
    • 3
  • Kenshi Hayashi
    • 2
  • Takuro Kobayashi
    • 1
  1. 1.Department of Neurology, Neurological Institute, Faculty of Medicine, Kyushu University 60, Fukuoka 812-82, Japan Fax: +81-92-642-5352; e-mail: furuya@neuro.med.kyushu-u.ac.jpJP
  2. 2.Division of Genome Analysis, Institute of Genetic Information, Kyushu University, Fukuoka, JapanJP
  3. 3.Division of Molecular Pathogenesis, Institute of Genetic Information, Kyushu University, Fukuoka, JapanJP
  4. 4.Neurological Division, Matsuyama Red Cross Hospital, Ehime, JapanJP
  5. 5.Department of Brain Pathophysiology, Faculty of Medicine, Kyoto University, Kyoto, JapanJP
  6. 6.Neurological Division, Iizuka General Hospital, Fukuoka, JapanJP
  7. 7.Department of Neurology, Neurological Institute, School of Medicine, Niigata University, Niigata, JapanJP

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