Identification and functional characterization of five novel mutant alleles in 58 Italian patients with Gaucher disease type 1.

Snjezana Miocić, Mirella Filocamo, Silvia Dominissini, Anna Lisa E Montalvo, Kristian Vlahovicek, Marta Deganuto, Raffaella Mazzotti, Roberta Cariati, Bruno Bembi, Maria Gabriela Pittis

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Abstract

Gaucher disease (GD) is the most frequent lysosomal glycolipid storage disorder due to an autosomal recessive deficiency of acid beta-glucosidase characterized by the accumulation of glucocerebroside. In this work we carried out the molecular analysis of the glucocerebrosidase gene (GBA) in 58 unrelated patients with GD type 1. We identified five novel genetic alterations: three missense changes c.187G>A (p.D63N), c.473T>G (p.I158S), c.689T>A (p.V230E), a gene-pseudogene recombinant allele and a non-pseudogene-derived complex allele [c.1379G>A;c.1469A>G] encoding [p.G460D;p.H490R]. All mutant alleles were present as compound heterozygotes in association with c.1226A>G (p.N409S), the most common mutation in GD1. The missense mutant proteins were expressed in vitro in COS-1 cells and analyzed by enzyme activity, protein processing and intracellular localization. Functional studies also included the c.662C>T (p.P221L) mutation recently reported in the Spanish GD population (Montfort et al., 2004). The missense mutant alleles retained an extremely low residual enzyme activity with respect to wild type; the complex allele expressed no activity. Processing of the mutant proteins was unaltered except for c.473T>G which was differently glycosylated due to the exposition of an additional glycosylation site. Immunofluorescence studies showed that protein trafficking into the lysosomes was unaffected in all cases. Finally, the characterization of the novel recombinant allele identified a crossover involving the GBA gene and pseudogene between intron 5 and exon 7. (c) 2004 Wiley-Liss, Inc.

Original languageEnglish
Pages (from-to)100
Number of pages1
JournalHuman Mutation
Volume25
Issue number1
Publication statusPublished - Jan 2005

Fingerprint

Gaucher Disease
Alleles
Glucosylceramidase
Pseudogenes
Mutant Proteins
Genes
Glucosylceramides
Mutation
COS Cells
Glycolipids
Protein Transport
Enzymes
Heterozygote
Lysosomes
Glycosylation
Introns
Fluorescent Antibody Technique
Exons
Population
Proteins

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Miocić, S., Filocamo, M., Dominissini, S., Montalvo, A. L. E., Vlahovicek, K., Deganuto, M., ... Pittis, M. G. (2005). Identification and functional characterization of five novel mutant alleles in 58 Italian patients with Gaucher disease type 1. Human Mutation, 25(1), 100.

Identification and functional characterization of five novel mutant alleles in 58 Italian patients with Gaucher disease type 1. / Miocić, Snjezana; Filocamo, Mirella; Dominissini, Silvia; Montalvo, Anna Lisa E; Vlahovicek, Kristian; Deganuto, Marta; Mazzotti, Raffaella; Cariati, Roberta; Bembi, Bruno; Pittis, Maria Gabriela.

In: Human Mutation, Vol. 25, No. 1, 01.2005, p. 100.

Research output: Contribution to journalArticle

Miocić, S, Filocamo, M, Dominissini, S, Montalvo, ALE, Vlahovicek, K, Deganuto, M, Mazzotti, R, Cariati, R, Bembi, B & Pittis, MG 2005, 'Identification and functional characterization of five novel mutant alleles in 58 Italian patients with Gaucher disease type 1.', Human Mutation, vol. 25, no. 1, pp. 100.
Miocić, Snjezana ; Filocamo, Mirella ; Dominissini, Silvia ; Montalvo, Anna Lisa E ; Vlahovicek, Kristian ; Deganuto, Marta ; Mazzotti, Raffaella ; Cariati, Roberta ; Bembi, Bruno ; Pittis, Maria Gabriela. / Identification and functional characterization of five novel mutant alleles in 58 Italian patients with Gaucher disease type 1. In: Human Mutation. 2005 ; Vol. 25, No. 1. pp. 100.
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