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Preserved proteinase K-resistant core after amplification of alpha-synuclein aggregates: Implication to disease-related structural study
- Source :
- Biochemical and Biophysical Research Communications. 522:655-661
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Many pathological proteins related to neurodegenerative diseases are misfolded, aggregating to form amyloid fibrils during pathogenesis. One of the pathological proteins, alpha-synuclein (α-syn), accumulates in the brains of Parkinson disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy (MSA), which are designated as synucleinopathies. Recently, structural properties of abnormal accumulated proteins are suggested to determine the disease phenotype. However, the biochemical and structural characteristics of those accumulated proteins are still poorly understood. We previously reported the sequence and seed-structure-dependent polymorphic fibrils of α-syn and the polymorphism was identified by proteinase K-resistant cores determined by mass spectrometry (MS) analysis. In this study, we applied this method to analyze α-syn aggregates of MSA and DLB. To perform MS analysis on proteinase K-resistant cores, we first performed amplification of α-syn aggregates by seeding reaction and protein misfolding cyclic amplification (PMCA) to obtain a sufficient amount of aggregates. Using SDS insoluble fraction of the disease brain, we successfully amplified enough α-syn aggregates for MS analysis. We differentiated between mouse and human α-syn aggregates by MS analysis on proteinase K-resistant cores of the aggregates before and after amplification. The results suggest that structural properties of amplified α-syn fibrils are preserved after PMCA and these methods can be applicable in the study of pathological proteins of the neurodegenerative disorders.
- Subjects :
- Male
0301 basic medicine
Synucleinopathies
Biophysics
Fibril
Protein Aggregation, Pathological
Biochemistry
Pathogenesis
Mice
Protein Aggregates
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Atrophy
medicine
Animals
Humans
Molecular Biology
Aged
Alpha-synuclein
biology
Dementia with Lewy bodies
Brain
Cell Biology
Middle Aged
medicine.disease
Proteinase K
nervous system diseases
030104 developmental biology
nervous system
chemistry
030220 oncology & carcinogenesis
alpha-Synuclein
biology.protein
Protein Misfolding Cyclic Amplification
Female
Endopeptidase K
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 522
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....41e996906bed476182270be4290c7a61