Abstract
Gaucher disease is the most frequent lysosomal storage disorder due to the deficiency of the acid β-glucosidase, encoded by the GBA gene. In this study, we report the structural and functional characterization of 11 novel GBA alleles. Seven single missense alleles, P159S, N188I, E235K, P245T, W312S, S366R and W381C, and two alleles carrying in cis mutations, (N188S; G265R) and (E326K; D380N), were studied for enzyme activity in transiently transfected cells. All mutants were inactive except the P159S, which retained 15% of wild-type activity. To further characterize the alleles carrying two in cis mutations, we expressed constructs bearing singly each mutation. The presence of G265R or D380N mutations completely abolished enzyme activity, while N188S and E326K mutants retained 25 and 54% of wild-type activity, respectively. Two mutations, affecting the acceptor splice site of introns 5 (c.589-1G>A) and 9 (c.1389-1G>A), led to the synthesis of aberrant mRNA. Unpredictably, family studies showed that two alleles resulted from germline or ‘de novo’ mutations. These results strengthen the importance of performing a complete and accurate molecular analysis of the GBA gene in order to avoid misleading conclusions and provide a comprehensive functional analysis of new GBA mutations.
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Acknowledgements
This work was supported by a grant of the Italian Ministry of Health PRF 37/08 ‘Clinical History and long-term cost-effectiveness of Enzyme Replacement Therapy (ERT) for Gaucher Disease in Italy.’ Some samples were obtained from the ‘Cell Line and DNA Biobank from Patients Affected by Genetic Diseases’ (G. Gaslini Institute) – Telethon Network of Genetic Biobanks (Project No. GTB12001A). We thank the Fondazione Pierfranco and Luisa Mariani of Milano for providing financial support for clinical assistance to the metabolic patients.
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Malini, E., Grossi, S., Deganuto, M. et al. Functional analysis of 11 novel GBA alleles. Eur J Hum Genet 22, 511–516 (2014). https://doi.org/10.1038/ejhg.2013.182
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DOI: https://doi.org/10.1038/ejhg.2013.182
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