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Proteomics-Based Approach Identifies Altered ER Domain Properties by ALS-Linked VAPB Mutation
- Source :
- Scientific Reports, Scientific Reports, Vol 10, Iss 1, Pp 1-16 (2020)
- Publication Year :
- 2019
-
Abstract
- An ER transmembrane protein, vesicle-associated membrane protein-associated protein B (VAPB), binds to several organelle-resident membrane proteins to mediate ER-organelle tethering. Mutation in amyotrophic lateral sclerosis (ALS) induces protein misfolding and aggregation, leading to ER disorganization. Gain or loss of function is suggested for VAPB mutation, however comprehensive study focusing on VAPB-ER domain has yet been performed. We here conducted proteomic characterization of the ER containing VAPB and its ALS-linked P56S mutant. For this purpose, we first optimized the proteomics of different ER domains immuno-isolated from cultured cells, and identified ER sheet- and tubule-specific proteomes. By using these as references, we found that VAPB-ER proteome had intermediate ER domain properties but its tubular property was specifically decreased by its mutation. Biochemical, immunofluorescence and proximity ligation assays suggested this was mediated by delocalization of VAPB from ER tubules. The VAPB-ER proteomics further suggested reduced incorporation of multiple proteins located in different organelles, which was confirmed by proximity ligation assay. Taken together, our proteomics-based approach indicates altered ER domain properties and impaired ER-organelle tethering by VAPB mutation.
- Subjects :
- Proteomics
Mutant
Vesicular Transport Proteins
lcsh:Medicine
Proteomic analysis
Proximity ligation assay
medicine.disease_cause
Endoplasmic Reticulum
Article
Cell Line
Mice
Protein Domains
Protein Interaction Mapping
medicine
Animals
Motor neuron disease
lcsh:Science
Mutation
Multidisciplinary
Chemistry
lcsh:R
Amyotrophic Lateral Sclerosis
VAPB
Transmembrane protein
Cell biology
Membrane protein
Proteome
lcsh:Q
Protein aggregation
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 10
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Scientific reports
- Accession number :
- edsair.doi.dedup.....d3cff9875002b6411034b2a27e2583ab