Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology
Complex asparagine-linked glycosylation plays key roles in cellular functions, including cellular signaling, protein stability, and immune response. Previously, we characterized the appearance of a complex asparagine-linked glycosylated form of lysosome-associated membrane protein 1 (LAMP1) in the c...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/9/5082 |
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| author | Niamh X. Cawley Anna T. Lyons Daniel Abebe Rachel Luke Julia Yerger Rebecca Telese Christopher A. Wassif Joan E. Bailey-Wilson Forbes D. Porter |
| author_facet | Niamh X. Cawley Anna T. Lyons Daniel Abebe Rachel Luke Julia Yerger Rebecca Telese Christopher A. Wassif Joan E. Bailey-Wilson Forbes D. Porter |
| author_sort | Niamh X. Cawley |
| collection | DOAJ |
| description | Complex asparagine-linked glycosylation plays key roles in cellular functions, including cellular signaling, protein stability, and immune response. Previously, we characterized the appearance of a complex asparagine-linked glycosylated form of lysosome-associated membrane protein 1 (LAMP1) in the cerebellum of <i>Npc1<sup>−/−</sup></i> mice. This LAMP1 form was found on activated microglia, and its appearance correlated both spatially and temporally with cerebellar Purkinje neuron loss. To test the importance of complex asparagine-linked glycosylation in NPC1 pathology, we generated NPC1 knock-out mice deficient in MGAT5, a key Golgi-resident glycosyl transferase involved in complex asparagine-linked glycosylation. Our results show that <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice were smaller than <i>Mgat5<sup>+/+</sup>:Npc1<sup>−/−</sup></i> mice, and exhibited earlier NPC1 disease onset and reduced lifespan. Western blot and lectin binding analyses of cerebellar extracts confirmed the reduction in complex asparagine-linked glycosylation, and the absence of the hyper-glycosylated LAMP1 previously observed. Western blot analysis of cerebellar extracts demonstrated reduced calbindin staining in <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice compared to <i>Mgat5<sup>+/+</sup>:Npc1<sup>−/−</sup></i> mutant mice, and immunofluorescent staining of cerebellar sections indicated decreased levels of Purkinje neurons and increased astrogliosis in <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice. Our results suggest that reduced asparagine-linked glycosylation increases NPC1 disease severity in mice, and leads to the hypothesis that mutations in genes involved in asparagine-linked glycosylation may contribute to disease severity progression in individuals with NPC1. To examine this with respect to <i>MGAT5,</i> we analyzed 111 NPC1 patients for two <i>MGAT5</i> SNPs associated with multiple sclerosis; however, we did not identify an association with NPC1 phenotypic severity. |
| first_indexed | 2024-03-10T04:04:28Z |
| format | Article |
| id | doaj.art-8a8f34aa46244eb9aff43153a06e4e10 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:04:28Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8a8f34aa46244eb9aff43153a06e4e102023-11-23T08:26:46ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-01239508210.3390/ijms23095082Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 PathologyNiamh X. Cawley0Anna T. Lyons1Daniel Abebe2Rachel Luke3Julia Yerger4Rebecca Telese5Christopher A. Wassif6Joan E. Bailey-Wilson7Forbes D. Porter8Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USAResearch Animal Management Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USAComputational and Statistical Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Department of Health and Human Services, Baltimore, MD 21224, USADivision of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USAComplex asparagine-linked glycosylation plays key roles in cellular functions, including cellular signaling, protein stability, and immune response. Previously, we characterized the appearance of a complex asparagine-linked glycosylated form of lysosome-associated membrane protein 1 (LAMP1) in the cerebellum of <i>Npc1<sup>−/−</sup></i> mice. This LAMP1 form was found on activated microglia, and its appearance correlated both spatially and temporally with cerebellar Purkinje neuron loss. To test the importance of complex asparagine-linked glycosylation in NPC1 pathology, we generated NPC1 knock-out mice deficient in MGAT5, a key Golgi-resident glycosyl transferase involved in complex asparagine-linked glycosylation. Our results show that <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice were smaller than <i>Mgat5<sup>+/+</sup>:Npc1<sup>−/−</sup></i> mice, and exhibited earlier NPC1 disease onset and reduced lifespan. Western blot and lectin binding analyses of cerebellar extracts confirmed the reduction in complex asparagine-linked glycosylation, and the absence of the hyper-glycosylated LAMP1 previously observed. Western blot analysis of cerebellar extracts demonstrated reduced calbindin staining in <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice compared to <i>Mgat5<sup>+/+</sup>:Npc1<sup>−/−</sup></i> mutant mice, and immunofluorescent staining of cerebellar sections indicated decreased levels of Purkinje neurons and increased astrogliosis in <i>Mgat5<sup>−/−</sup>:Npc1<sup>−/−</sup></i> mice. Our results suggest that reduced asparagine-linked glycosylation increases NPC1 disease severity in mice, and leads to the hypothesis that mutations in genes involved in asparagine-linked glycosylation may contribute to disease severity progression in individuals with NPC1. To examine this with respect to <i>MGAT5,</i> we analyzed 111 NPC1 patients for two <i>MGAT5</i> SNPs associated with multiple sclerosis; however, we did not identify an association with NPC1 phenotypic severity.https://www.mdpi.com/1422-0067/23/9/5082Niemann–Pick diseasetype CN-linked glycosylationMGAT5LAMP1disease severity score |
| spellingShingle | Niamh X. Cawley Anna T. Lyons Daniel Abebe Rachel Luke Julia Yerger Rebecca Telese Christopher A. Wassif Joan E. Bailey-Wilson Forbes D. Porter Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology International Journal of Molecular Sciences Niemann–Pick disease type C N-linked glycosylation MGAT5 LAMP1 disease severity score |
| title | Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology |
| title_full | Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology |
| title_fullStr | Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology |
| title_full_unstemmed | Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology |
| title_short | Complex N-Linked Glycosylation: A Potential Modifier of Niemann–Pick Disease, Type C1 Pathology |
| title_sort | complex n linked glycosylation a potential modifier of niemann pick disease type c1 pathology |
| topic | Niemann–Pick disease type C N-linked glycosylation MGAT5 LAMP1 disease severity score |
| url | https://www.mdpi.com/1422-0067/23/9/5082 |
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