Molecular characterization of mutations that cause globoid cell leukodystrophy and pharmacological rescue using small molecule chemical chaperones

Wing C. Lee, Dongcheul Kang, Ena Causevic, Aimee R. Herdt, Elizabeth A. Eckman, Christopher B. Eckman

Research output: Contribution to journalArticle

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Abstract

Globoid cell leukodystrophy (GLD) (Krabbe disease) is an autosomal recessive, degenerative, lysosomal storage disease caused by a severe loss of galactocerebrosidase (GALC) enzymatic activity. Of the >70 disease-causing mutations in the GALC gene, most are located outside of the catalytic domain of the enzyme. To determine how GALC mutations impair enzymatic activity, we investigated the impact of multiple disease-causing mutations on GALC processing, localization, and enzymatic activity. Studies in mammalian cells revealed dramatic decreases in GALC activity and a lack of appropriate protein processing into an N-terminal GALC fragment for each of the mutants examined. Consistent with this, we observed significantly less GALC localized to the lysosome and impairment in either the secretion or reuptake of mutant GALC. Notably, the D528N mutation was found to induce hyperglycosylation and protein misfolding. Reversal of these conditions resulted in an increase in proper processing and GALC activity, suggesting that glycosylation may play a critical role in the disease process in patients with this mutation. Recent studies have shown that enzyme inhibitors can sometimes "chaperone" misfolded polypeptides to their appropriate target organelle, bypassing the normal cellular quality control machinery and resulting in enhanced activity. To determine whether this may also work for GLD, we examined the effect of α-lobeline, an inhibitor of GALC, on D528N mutant cells. After treatment,GALCactivity was significantly increased. This study suggests that mutations in GALC can cause GLD by impairing protein processing and/or folding and that pharmacological chaperones may be potential therapeutic agents for patients carrying certain mutations. Copyright

Original languageEnglish (US)
Pages (from-to)5489-5497
Number of pages9
JournalJournal of Neuroscience
Volume30
Issue number16
DOIs
StatePublished - Apr 21 2010

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Galactosylceramidase
Globoid Cell Leukodystrophy
Pharmacology
Mutation
Lobeline
Lysosomal Storage Diseases
Proteins
Proxy
Enzyme Inhibitors
Lysosomes
Glycosylation
Quality Control
Organelles

ASJC Scopus subject areas

  • Neuroscience(all)

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Molecular characterization of mutations that cause globoid cell leukodystrophy and pharmacological rescue using small molecule chemical chaperones. / Lee, Wing C.; Kang, Dongcheul; Causevic, Ena; Herdt, Aimee R.; Eckman, Elizabeth A.; Eckman, Christopher B.

In: Journal of Neuroscience, Vol. 30, No. 16, 21.04.2010, p. 5489-5497.

Research output: Contribution to journalArticle

Lee, Wing C. ; Kang, Dongcheul ; Causevic, Ena ; Herdt, Aimee R. ; Eckman, Elizabeth A. ; Eckman, Christopher B. / Molecular characterization of mutations that cause globoid cell leukodystrophy and pharmacological rescue using small molecule chemical chaperones. In: Journal of Neuroscience. 2010 ; Vol. 30, No. 16. pp. 5489-5497.
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