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N-glycome of the lysosomal glycocalyx is altered in Niemann-Pick Type C disease model cells

Košiček, Marko; Gudelj, Ivan; Horvatić, Anita; Jović, Tanja; Vučković, Frano; Lauc, Gordan; Hećimović, Silva (2018) N-glycome of the lysosomal glycocalyx is altered in Niemann-Pick Type C disease model cells. Molecular & cellular proteomics, 17 (4). pp. 631-642. ISSN 1535-9476

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Increasing evidence implicates lysosomal dysfunction in the pathogenesis of neurodegenerative diseases, including the rare inherited lysosomal storage disorders (LSDs) and the most common neurodegenerative diseases, such as Alzheimer's and Parkinson's disease (AD and PD). Although the triggers of the lysosomal impairment may involve the accumulated macromolecules or dysfunction of the lysosomal enzymes, the role of the lysosomal glycocalyx in the lysosomal (dys)function has not been studied. The goal of this work was to analyze whether there are changes in the lysosomal glycocalyx in a cellular model of a LSD Niemann-Pick type C disease (NPC). Using the ferrofluid nanoparticles we isolated lysosomal organelles from NPC1-null and CHOwt cells. The magnetically isolated lysosomal fractions were enriched with the lysosomal marker protein LAMP1 and showed the key features of NPC disease: 3-fold higher cholesterol content and 4–5 fold enlarged size of the particles compared with the lysosomal fractions of wt cells. These lysosomal fractions were further processed to isolate lysosomal membrane proteins using Triton X-114 and their N- glycome was analyzed by HILIC-UPLC. N-glycans presented in each chromatographic peak were elucidated using MALDI-TOF/TOF-MS. We detected changes in the N-glycosylation pattern of the lysosomal glycocalyx of NPC1-null versus wt cells which involved high-mannose and sialylated N-glycans. To the best of our knowledge this study is the first to report N- glycome profiling of the lysosomal glycocalyx in NPC disease cellular model and the first to report the specific changes in the lysosomal glycocalyx in NPC1-null cells. We speculate that changes in the lysosomal glycocalyx may contribute to lysosomal (dys)function. Further glycome profiling of the lysosomal glycocalyx in other LSDs as well as the most common neurodegenerative diseases, such as AD and PD, is necessary to better understand the role of the lysosomal glycocalyx and to reveal its potential contribution in lysosomal dysfunction leading to neurodegeneration.

Item Type: Article
Additional Information: This work was funded by the Unity Through Knowledge Fund (S.H.), by the European Commission FP7 grants MIMOmics (contract #305280, G.L.), HTP-GlycoMet (contract #324400, G.L.), VetMedZg (contract #621394, A.H.), H2020 grants GlySign (contract #722095, G.L.), SYSCID (contract #733100, G.L.) and IMforFuture (contract #721815, G.L.) as well as by funding for the Croatian National Centre of Research Excellence in Personalized Healthcare and IRI project Nova generacija visokoprotocnih glikoservisa.
Uncontrolled Keywords: Niemann-Pick Type C disease ; lysosomal dysfunction ; lysosomal glycocalyx ; neurodegeneration ; glycomics ; proteomics
Subjects: NATURAL SCIENCES > Biology
Divisions: Division of Molecular Medicine
Project titleProject leaderProject codeProject type
Methods for Integrated analysis of Multiple Omics datasets-MIMOMICSUNSPECIFIED305280EK
Methods for high-throughput glycoproteomic analysis-HTP-GLYCOMETUNSPECIFIED324400EK
Upgrading the research performance in molecular medicine at the Faculty of Veterinary Medicine University of Zagreb-VETMEDZGUNSPECIFIED621394EK
Exploitation of Glycosylation Signatures for Precision Medicine-GlySignUNSPECIFIED722095EK
A Systems medicine approach to chronic inflammatory disease-SYSCIDUNSPECIFIED733100EK
Innovative training in methods for future data-IMforFUTUREUNSPECIFIED721815EK
Depositing User: Dr. sc. Silva Katušić Hećimović
Date Deposited: 28 Nov 2019 11:19
DOI: 10.1074/mcp.RA117.000129

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