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Stabilization of α-synuclein oligomers using formaldehyde

The group of neurodegenerative diseases, Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) all exhibit inclusions containing amyloid-type α-synuclein (α-syn) aggregates within degenerating brain cells. α-syn also exists as soluble oligomeric species th...

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Autores principales: Ruesink, Harm, Reimer, Lasse, Gregersen, Emil, Moeller, Arne, Betzer, Cristine, Jensen, Poul Henning
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6788717/
https://www.ncbi.nlm.nih.gov/pubmed/31603909
http://dx.doi.org/10.1371/journal.pone.0216764
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author Ruesink, Harm
Reimer, Lasse
Gregersen, Emil
Moeller, Arne
Betzer, Cristine
Jensen, Poul Henning
author_facet Ruesink, Harm
Reimer, Lasse
Gregersen, Emil
Moeller, Arne
Betzer, Cristine
Jensen, Poul Henning
author_sort Ruesink, Harm
collection PubMed
description The group of neurodegenerative diseases, Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) all exhibit inclusions containing amyloid-type α-synuclein (α-syn) aggregates within degenerating brain cells. α-syn also exists as soluble oligomeric species that are hypothesized to represent intermediates between its native and aggregated states. These oligomers are present in brain extracts from patients suffering from synucleinopathies and hold great potential as biomarkers. Although easily prepared in vitro, oligomers are metastable and dissociate over time, thereby complicating α-syn oligomer research. Using the small amine-reactive cross-linker, formaldehyde (FA), we successfully stabilized α-syn oligomers without affecting their size, overall structure or antigenicity towards aggregate-conformation specific α-syn antibodies FILA and MJFR-14-6-4-2. Further, cross-linked α-syn oligomers show resistance towards denaturant like urea and SDS treatment and remain fully functional as internal standard in an aggregation-specific enzyme-linked immunosorbent assay (ELISA) despite prior incubation with urea. We propose that FA cross-linked α-syn oligomers could serve as important calibrators to facilitate comparative and standardized α-syn biomarker studies going forward.
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spelling pubmed-67887172019-10-25 Stabilization of α-synuclein oligomers using formaldehyde Ruesink, Harm Reimer, Lasse Gregersen, Emil Moeller, Arne Betzer, Cristine Jensen, Poul Henning PLoS One Research Article The group of neurodegenerative diseases, Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) all exhibit inclusions containing amyloid-type α-synuclein (α-syn) aggregates within degenerating brain cells. α-syn also exists as soluble oligomeric species that are hypothesized to represent intermediates between its native and aggregated states. These oligomers are present in brain extracts from patients suffering from synucleinopathies and hold great potential as biomarkers. Although easily prepared in vitro, oligomers are metastable and dissociate over time, thereby complicating α-syn oligomer research. Using the small amine-reactive cross-linker, formaldehyde (FA), we successfully stabilized α-syn oligomers without affecting their size, overall structure or antigenicity towards aggregate-conformation specific α-syn antibodies FILA and MJFR-14-6-4-2. Further, cross-linked α-syn oligomers show resistance towards denaturant like urea and SDS treatment and remain fully functional as internal standard in an aggregation-specific enzyme-linked immunosorbent assay (ELISA) despite prior incubation with urea. We propose that FA cross-linked α-syn oligomers could serve as important calibrators to facilitate comparative and standardized α-syn biomarker studies going forward. Public Library of Science 2019-10-11 /pmc/articles/PMC6788717/ /pubmed/31603909 http://dx.doi.org/10.1371/journal.pone.0216764 Text en © 2019 Ruesink et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Ruesink, Harm
Reimer, Lasse
Gregersen, Emil
Moeller, Arne
Betzer, Cristine
Jensen, Poul Henning
Stabilization of α-synuclein oligomers using formaldehyde
title Stabilization of α-synuclein oligomers using formaldehyde
title_full Stabilization of α-synuclein oligomers using formaldehyde
title_fullStr Stabilization of α-synuclein oligomers using formaldehyde
title_full_unstemmed Stabilization of α-synuclein oligomers using formaldehyde
title_short Stabilization of α-synuclein oligomers using formaldehyde
title_sort stabilization of α-synuclein oligomers using formaldehyde
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6788717/
https://www.ncbi.nlm.nih.gov/pubmed/31603909
http://dx.doi.org/10.1371/journal.pone.0216764
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