Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates

Goki Tanaka, Tomoyuki Yamanaka, Yoshiaki Furukawa, Naoko Kajimura, Kaoru Mitsuoka, Nobuyuki Nukina

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Alpha-synuclein (a-syn) aggregation in brain is implicated in several synucleinopathies, including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Until date, at least six disease-associated mutations in a-syn (namely A30P, E46K, H50Q, G51D, A53T, and A53E) are known to cause dominantly inherited familial forms of synucleinopathies. Previous studies using recombinant proteins have reported that a subset of disease-associated mutants show higher aggregation propensities and form spectroscopically distinguishable aggregates compared to wild-type (WT). However, morphological and biochemical comparison of the aggregates for all disease-associated a-syn mutants have not yet been performed. In this study, we performed electron microscopic examination, guanidinium hydrochloride (GdnHCl) denaturation, and protease digestion to classify the aggregates from their respective point mutations. Using electron microscopy we observed variations of amyloid fibrillar morphologies among the aggregates of a-syn mutants, mainly categorized into two groups: twisted fibrils observed for both WT and E46K while straight fibrils for the other mutants. GdnHCl denaturation experiments revealed the a-syn mutants except for E46K were more resistant than WT against the denaturation. Mass spectrometry analysis of protease-treated aggregates showed a variety of protease-resistant cores, which may correspond to their morphological properties. The difference of their properties could be implicated in the clinicopathological difference of synucleinopathies with those mutations.

Original languageEnglish
Pages (from-to)729-734
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume508
Issue number3
DOIs
Publication statusPublished - 2019 Jan 15

Fingerprint

alpha-Synuclein
Peptide Hydrolases
Guanidine
Denaturation
Multiple System Atrophy
Lewy Body Disease
Mutation
Point Mutation
Recombinant Proteins
Amyloid
Agglomeration
Parkinson Disease
Digestion
Mass Spectrometry
Electron Microscopy
Electrons
Electron microscopy
Brain
Mass spectrometry
Microscopic examination

Keywords

  • Alpha-synuclein
  • Disease-associated mutation
  • Protein aggregation
  • Synucleinopathy

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates. / Tanaka, Goki; Yamanaka, Tomoyuki; Furukawa, Yoshiaki; Kajimura, Naoko; Mitsuoka, Kaoru; Nukina, Nobuyuki.

In: Biochemical and Biophysical Research Communications, Vol. 508, No. 3, 15.01.2019, p. 729-734.

Research output: Contribution to journalArticle

Tanaka, Goki ; Yamanaka, Tomoyuki ; Furukawa, Yoshiaki ; Kajimura, Naoko ; Mitsuoka, Kaoru ; Nukina, Nobuyuki. / Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates. In: Biochemical and Biophysical Research Communications. 2019 ; Vol. 508, No. 3. pp. 729-734.
@article{0df95dfea3074783a796aa8f67f0a112,
title = "Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates",
abstract = "Alpha-synuclein (a-syn) aggregation in brain is implicated in several synucleinopathies, including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Until date, at least six disease-associated mutations in a-syn (namely A30P, E46K, H50Q, G51D, A53T, and A53E) are known to cause dominantly inherited familial forms of synucleinopathies. Previous studies using recombinant proteins have reported that a subset of disease-associated mutants show higher aggregation propensities and form spectroscopically distinguishable aggregates compared to wild-type (WT). However, morphological and biochemical comparison of the aggregates for all disease-associated a-syn mutants have not yet been performed. In this study, we performed electron microscopic examination, guanidinium hydrochloride (GdnHCl) denaturation, and protease digestion to classify the aggregates from their respective point mutations. Using electron microscopy we observed variations of amyloid fibrillar morphologies among the aggregates of a-syn mutants, mainly categorized into two groups: twisted fibrils observed for both WT and E46K while straight fibrils for the other mutants. GdnHCl denaturation experiments revealed the a-syn mutants except for E46K were more resistant than WT against the denaturation. Mass spectrometry analysis of protease-treated aggregates showed a variety of protease-resistant cores, which may correspond to their morphological properties. The difference of their properties could be implicated in the clinicopathological difference of synucleinopathies with those mutations.",
keywords = "Alpha-synuclein, Disease-associated mutation, Protein aggregation, Synucleinopathy",
author = "Goki Tanaka and Tomoyuki Yamanaka and Yoshiaki Furukawa and Naoko Kajimura and Kaoru Mitsuoka and Nobuyuki Nukina",
year = "2019",
month = "1",
day = "15",
doi = "10.1016/j.bbrc.2018.11.200",
language = "English",
volume = "508",
pages = "729--734",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates

AU - Tanaka, Goki

AU - Yamanaka, Tomoyuki

AU - Furukawa, Yoshiaki

AU - Kajimura, Naoko

AU - Mitsuoka, Kaoru

AU - Nukina, Nobuyuki

PY - 2019/1/15

Y1 - 2019/1/15

N2 - Alpha-synuclein (a-syn) aggregation in brain is implicated in several synucleinopathies, including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Until date, at least six disease-associated mutations in a-syn (namely A30P, E46K, H50Q, G51D, A53T, and A53E) are known to cause dominantly inherited familial forms of synucleinopathies. Previous studies using recombinant proteins have reported that a subset of disease-associated mutants show higher aggregation propensities and form spectroscopically distinguishable aggregates compared to wild-type (WT). However, morphological and biochemical comparison of the aggregates for all disease-associated a-syn mutants have not yet been performed. In this study, we performed electron microscopic examination, guanidinium hydrochloride (GdnHCl) denaturation, and protease digestion to classify the aggregates from their respective point mutations. Using electron microscopy we observed variations of amyloid fibrillar morphologies among the aggregates of a-syn mutants, mainly categorized into two groups: twisted fibrils observed for both WT and E46K while straight fibrils for the other mutants. GdnHCl denaturation experiments revealed the a-syn mutants except for E46K were more resistant than WT against the denaturation. Mass spectrometry analysis of protease-treated aggregates showed a variety of protease-resistant cores, which may correspond to their morphological properties. The difference of their properties could be implicated in the clinicopathological difference of synucleinopathies with those mutations.

AB - Alpha-synuclein (a-syn) aggregation in brain is implicated in several synucleinopathies, including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Until date, at least six disease-associated mutations in a-syn (namely A30P, E46K, H50Q, G51D, A53T, and A53E) are known to cause dominantly inherited familial forms of synucleinopathies. Previous studies using recombinant proteins have reported that a subset of disease-associated mutants show higher aggregation propensities and form spectroscopically distinguishable aggregates compared to wild-type (WT). However, morphological and biochemical comparison of the aggregates for all disease-associated a-syn mutants have not yet been performed. In this study, we performed electron microscopic examination, guanidinium hydrochloride (GdnHCl) denaturation, and protease digestion to classify the aggregates from their respective point mutations. Using electron microscopy we observed variations of amyloid fibrillar morphologies among the aggregates of a-syn mutants, mainly categorized into two groups: twisted fibrils observed for both WT and E46K while straight fibrils for the other mutants. GdnHCl denaturation experiments revealed the a-syn mutants except for E46K were more resistant than WT against the denaturation. Mass spectrometry analysis of protease-treated aggregates showed a variety of protease-resistant cores, which may correspond to their morphological properties. The difference of their properties could be implicated in the clinicopathological difference of synucleinopathies with those mutations.

KW - Alpha-synuclein

KW - Disease-associated mutation

KW - Protein aggregation

KW - Synucleinopathy

UR - http://www.scopus.com/inward/record.url?scp=85059243329&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85059243329&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2018.11.200

DO - 10.1016/j.bbrc.2018.11.200

M3 - Article

C2 - 30528390

AN - SCOPUS:85059243329

VL - 508

SP - 729

EP - 734

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 3

ER -