Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells.
Sandhoff and Tay-Sachs disease are autosomal recessive GM2 gangliosidoses where a deficiency of lysosomal beta-hexosaminidase results in storage of glycoconjugates. Imino sugar (2-acetamido-1,4-imino-1,2,4-trideoxy-L-arabinitol) inhibition of beta-hexosaminidase in murine RAW264.7 macrophage-like ce...
| Auteurs principaux: | , , , , , |
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| Format: | Journal article |
| Langue: | English |
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2010
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| _version_ | 1826301607310000128 |
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| author | Boomkamp, S Rountree, J Neville, D Dwek, R Fleet, G Butters, T |
| author_facet | Boomkamp, S Rountree, J Neville, D Dwek, R Fleet, G Butters, T |
| author_sort | Boomkamp, S |
| collection | OXFORD |
| description | Sandhoff and Tay-Sachs disease are autosomal recessive GM2 gangliosidoses where a deficiency of lysosomal beta-hexosaminidase results in storage of glycoconjugates. Imino sugar (2-acetamido-1,4-imino-1,2,4-trideoxy-L-arabinitol) inhibition of beta-hexosaminidase in murine RAW264.7 macrophage-like cells led to lysosomal storage of glycoconjugates that were characterised structurally using fluorescence labelling of the free or glycolipid-derived oligosaccharides followed by HPLC and mass spectrometry. Stored glycoconjugates were confirmed as containing non-reducing GlcNAc or GalNAc residues resulting from the incomplete degradation of N-linked glycoprotein oligosaccharide and glycolipids, respectively. When substrate reduction therapeutics N-butyl-deoxynojirimycin (NB-DNJ) or N-butyldeoxygalactonojirimycin (NB-DGJ) were applied to the storage phenotype cells, an increase in glucosylated and galactosylated oligosaccharide species was observed due to endoplasmic reticulum alpha-glucosidases and lysosomal beta-galactosidase inhibition, respectively. Hexosaminidase inhibition triggered a tightly regulated cytokine-mediated inflammatory response that was normalised using imino sugars NB-DNJ and NB-DGJ, which restored the GM2 ganglioside storage burden but failed to reduce the levels of GA2 glycolipid or glycoprotein-derived N-linked oligosaccharides. Using a chemically induced gangliosidosis phenotype that can be modulated with substrate lowering drugs, the critical role of GM2 ganglioside in the progression of inflammatory disease is also demonstrated. |
| first_indexed | 2024-03-07T05:34:57Z |
| format | Journal article |
| id | oxford-uuid:e39483a4-2d81-4ffc-8170-438a5d3d2bea |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:34:57Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | oxford-uuid:e39483a4-2d81-4ffc-8170-438a5d3d2bea2022-03-27T10:09:59ZLysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:e39483a4-2d81-4ffc-8170-438a5d3d2beaEnglishSymplectic Elements at Oxford2010Boomkamp, SRountree, JNeville, DDwek, RFleet, GButters, TSandhoff and Tay-Sachs disease are autosomal recessive GM2 gangliosidoses where a deficiency of lysosomal beta-hexosaminidase results in storage of glycoconjugates. Imino sugar (2-acetamido-1,4-imino-1,2,4-trideoxy-L-arabinitol) inhibition of beta-hexosaminidase in murine RAW264.7 macrophage-like cells led to lysosomal storage of glycoconjugates that were characterised structurally using fluorescence labelling of the free or glycolipid-derived oligosaccharides followed by HPLC and mass spectrometry. Stored glycoconjugates were confirmed as containing non-reducing GlcNAc or GalNAc residues resulting from the incomplete degradation of N-linked glycoprotein oligosaccharide and glycolipids, respectively. When substrate reduction therapeutics N-butyl-deoxynojirimycin (NB-DNJ) or N-butyldeoxygalactonojirimycin (NB-DGJ) were applied to the storage phenotype cells, an increase in glucosylated and galactosylated oligosaccharide species was observed due to endoplasmic reticulum alpha-glucosidases and lysosomal beta-galactosidase inhibition, respectively. Hexosaminidase inhibition triggered a tightly regulated cytokine-mediated inflammatory response that was normalised using imino sugars NB-DNJ and NB-DGJ, which restored the GM2 ganglioside storage burden but failed to reduce the levels of GA2 glycolipid or glycoprotein-derived N-linked oligosaccharides. Using a chemically induced gangliosidosis phenotype that can be modulated with substrate lowering drugs, the critical role of GM2 ganglioside in the progression of inflammatory disease is also demonstrated. |
| spellingShingle | Boomkamp, S Rountree, J Neville, D Dwek, R Fleet, G Butters, T Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title | Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title_full | Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title_fullStr | Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title_full_unstemmed | Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title_short | Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells. |
| title_sort | lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells |
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