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High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors

An increasing number of neurodegenerative diseases are being found to be associated with the abnormal accumulation of aggregated proteins in the brain. In Parkinson’s disease, this process involves the aggregation of alpha-synuclein (α-syn) into intraneuronal inclusions. Thus, compounds that inhibit...

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Autores principales: Pujols, Jordi, Peña-Díaz, Samuel, Conde-Giménez, María, Pinheiro, Francisca, Navarro, Susanna, Sancho, Javier, Ventura, Salvador
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5372494/
https://www.ncbi.nlm.nih.gov/pubmed/28257086
http://dx.doi.org/10.3390/ijms18030478
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author Pujols, Jordi
Peña-Díaz, Samuel
Conde-Giménez, María
Pinheiro, Francisca
Navarro, Susanna
Sancho, Javier
Ventura, Salvador
author_facet Pujols, Jordi
Peña-Díaz, Samuel
Conde-Giménez, María
Pinheiro, Francisca
Navarro, Susanna
Sancho, Javier
Ventura, Salvador
author_sort Pujols, Jordi
collection PubMed
description An increasing number of neurodegenerative diseases are being found to be associated with the abnormal accumulation of aggregated proteins in the brain. In Parkinson’s disease, this process involves the aggregation of alpha-synuclein (α-syn) into intraneuronal inclusions. Thus, compounds that inhibit α-syn aggregation represent a promising therapeutic strategy as disease-modifying agents for neurodegeneration. The formation of α-syn amyloid aggregates can be reproduced in vitro by incubation of the recombinant protein. However, the in vitro aggregation of α-syn is exceedingly slow and highly irreproducible, therefore precluding fast high throughput anti-aggregation drug screening. Here, we present a simple and easy-to-implement in-plate method for screening large chemical libraries in the search for α-syn aggregation modulators. It allows us to monitor aggregation kinetics with high reproducibility, while being faster and requiring lower protein amounts than conventional aggregation assays. We illustrate how the approach enables the identification of strong aggregation inhibitors in a library of more than 14,000 compounds.
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spelling pubmed-53724942017-04-10 High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors Pujols, Jordi Peña-Díaz, Samuel Conde-Giménez, María Pinheiro, Francisca Navarro, Susanna Sancho, Javier Ventura, Salvador Int J Mol Sci Article An increasing number of neurodegenerative diseases are being found to be associated with the abnormal accumulation of aggregated proteins in the brain. In Parkinson’s disease, this process involves the aggregation of alpha-synuclein (α-syn) into intraneuronal inclusions. Thus, compounds that inhibit α-syn aggregation represent a promising therapeutic strategy as disease-modifying agents for neurodegeneration. The formation of α-syn amyloid aggregates can be reproduced in vitro by incubation of the recombinant protein. However, the in vitro aggregation of α-syn is exceedingly slow and highly irreproducible, therefore precluding fast high throughput anti-aggregation drug screening. Here, we present a simple and easy-to-implement in-plate method for screening large chemical libraries in the search for α-syn aggregation modulators. It allows us to monitor aggregation kinetics with high reproducibility, while being faster and requiring lower protein amounts than conventional aggregation assays. We illustrate how the approach enables the identification of strong aggregation inhibitors in a library of more than 14,000 compounds. MDPI 2017-03-02 /pmc/articles/PMC5372494/ /pubmed/28257086 http://dx.doi.org/10.3390/ijms18030478 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pujols, Jordi
Peña-Díaz, Samuel
Conde-Giménez, María
Pinheiro, Francisca
Navarro, Susanna
Sancho, Javier
Ventura, Salvador
High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title_full High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title_fullStr High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title_full_unstemmed High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title_short High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors
title_sort high-throughput screening methodology to identify alpha-synuclein aggregation inhibitors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5372494/
https://www.ncbi.nlm.nih.gov/pubmed/28257086
http://dx.doi.org/10.3390/ijms18030478
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