Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia
The GBA2 gene encodes the non-lysosomal glucosylceramidase (NLGase), an enzyme that catalyzes the conversion of glucosylceramide (GlcCer) to ceramide and glucose. Mutations in GBA2 have been associated with the development of neurological disorders such as autosomal recessive cerebellar ataxia, here...
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doaj-4c10326ea8924aeaa8c2371324a735472020-11-24T21:24:58ZengMDPI AGInternational Journal of Molecular Sciences1422-00672018-10-011910309910.3390/ijms19103099ijms19103099Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic AtaxiaAnna Malekkou0Maura Samarani1Anthi Drousiotou2Christina Votsi3Sandro Sonnino4Marios Pantzaris5Elena Chiricozzi6Eleni Zamba-Papanicolaou7Massimo Aureli8Nicoletta Loberto9Kyproula Christodoulou10Biochemical Genetics Department, The Cyprus Institute of Neurology and Genetics, Nicosia 1683, CyprusDepartment of Medical Biotechnology and Translational Medicine, University of Milano, 20122 Milano, ItalyBiochemical Genetics Department, The Cyprus Institute of Neurology and Genetics, Nicosia 1683, CyprusCyprus School of Molecular Medicine, Nicosia 1683, CyprusDepartment of Medical Biotechnology and Translational Medicine, University of Milano, 20122 Milano, ItalyCyprus School of Molecular Medicine, Nicosia 1683, CyprusDepartment of Medical Biotechnology and Translational Medicine, University of Milano, 20122 Milano, ItalyCyprus School of Molecular Medicine, Nicosia 1683, CyprusDepartment of Medical Biotechnology and Translational Medicine, University of Milano, 20122 Milano, ItalyDepartment of Medical Biotechnology and Translational Medicine, University of Milano, 20122 Milano, ItalyCyprus School of Molecular Medicine, Nicosia 1683, CyprusThe GBA2 gene encodes the non-lysosomal glucosylceramidase (NLGase), an enzyme that catalyzes the conversion of glucosylceramide (GlcCer) to ceramide and glucose. Mutations in GBA2 have been associated with the development of neurological disorders such as autosomal recessive cerebellar ataxia, hereditary spastic paraplegia, and Marinesco-Sjogren-Like Syndrome. Our group has previously identified the GBA2 c.1780G>C [p.Asp594His] missense mutation, in a Cypriot consanguineous family with spastic ataxia. In this study, we carried out a biochemical characterization of lymphoblastoid cell lines (LCLs) derived from three patients of this family. We found that the mutation strongly reduce NLGase activity both intracellularly and at the plasma membrane level. Additionally, we observed a two-fold increase of GlcCer content in LCLs derived from patients compared to controls, with the C16 lipid being the most abundant GlcCer species. Moreover, we showed that there is an apparent compensatory effect between NLGase and the lysosomal glucosylceramidase (GCase), since we found that the activity of GCase was three-fold higher in LCLs derived from patients compared to controls. We conclude that the c.1780G>C mutation results in NLGase loss of function with abolishment of the enzymatic activity and accumulation of GlcCer accompanied by a compensatory increase in GCase.http://www.mdpi.com/1422-0067/19/10/3099GBA2non-lysosomal β-glucosylceramidaseβ-glucocerebrosidasespastic ataxiaglucosylceramideplasma membranelymphoblastoid cell lines |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anna Malekkou Maura Samarani Anthi Drousiotou Christina Votsi Sandro Sonnino Marios Pantzaris Elena Chiricozzi Eleni Zamba-Papanicolaou Massimo Aureli Nicoletta Loberto Kyproula Christodoulou |
spellingShingle |
Anna Malekkou Maura Samarani Anthi Drousiotou Christina Votsi Sandro Sonnino Marios Pantzaris Elena Chiricozzi Eleni Zamba-Papanicolaou Massimo Aureli Nicoletta Loberto Kyproula Christodoulou Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia International Journal of Molecular Sciences GBA2 non-lysosomal β-glucosylceramidase β-glucocerebrosidase spastic ataxia glucosylceramide plasma membrane lymphoblastoid cell lines |
author_facet |
Anna Malekkou Maura Samarani Anthi Drousiotou Christina Votsi Sandro Sonnino Marios Pantzaris Elena Chiricozzi Eleni Zamba-Papanicolaou Massimo Aureli Nicoletta Loberto Kyproula Christodoulou |
author_sort |
Anna Malekkou |
title |
Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia |
title_short |
Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia |
title_full |
Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia |
title_fullStr |
Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia |
title_full_unstemmed |
Biochemical Characterization of the GBA2 c.1780G>C Missense Mutation in Lymphoblastoid Cells from Patients with Spastic Ataxia |
title_sort |
biochemical characterization of the gba2 c.1780g>c missense mutation in lymphoblastoid cells from patients with spastic ataxia |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2018-10-01 |
description |
The GBA2 gene encodes the non-lysosomal glucosylceramidase (NLGase), an enzyme that catalyzes the conversion of glucosylceramide (GlcCer) to ceramide and glucose. Mutations in GBA2 have been associated with the development of neurological disorders such as autosomal recessive cerebellar ataxia, hereditary spastic paraplegia, and Marinesco-Sjogren-Like Syndrome. Our group has previously identified the GBA2 c.1780G>C [p.Asp594His] missense mutation, in a Cypriot consanguineous family with spastic ataxia. In this study, we carried out a biochemical characterization of lymphoblastoid cell lines (LCLs) derived from three patients of this family. We found that the mutation strongly reduce NLGase activity both intracellularly and at the plasma membrane level. Additionally, we observed a two-fold increase of GlcCer content in LCLs derived from patients compared to controls, with the C16 lipid being the most abundant GlcCer species. Moreover, we showed that there is an apparent compensatory effect between NLGase and the lysosomal glucosylceramidase (GCase), since we found that the activity of GCase was three-fold higher in LCLs derived from patients compared to controls. We conclude that the c.1780G>C mutation results in NLGase loss of function with abolishment of the enzymatic activity and accumulation of GlcCer accompanied by a compensatory increase in GCase. |
topic |
GBA2 non-lysosomal β-glucosylceramidase β-glucocerebrosidase spastic ataxia glucosylceramide plasma membrane lymphoblastoid cell lines |
url |
http://www.mdpi.com/1422-0067/19/10/3099 |
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