title: High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Autonomous University of Barcelona, 08193 Bellaterra, Spain. jordi.pujolspujol@gmail.com.
authors.forename: Jordi
authors.initials: J
authors.lastname: Pujols
authors.affiliation: Department of Biochemistry and Molecular Biology, Autonomous University of Barcelona, 08193 Bellaterra, Spain. samuel.pdiaz@gmail.com.
authors.forename: Samuel
authors.initials: S
authors.lastname: Peña-Díaz
authors.affiliation: Department of Biochemistry and Molecular and Cell Biology, Institute for Biocomputation and Physics of Complex Systems (BIFI), University of Zaragoza, 50018 Zaragoza, Spain. maria.conde@gmail.com.
authors.forename: María
authors.initials: M
authors.lastname: Conde-Giménez
authors.affiliation: Department of Biochemistry and Molecular Biology, Autonomous University of Barcelona, 08193 Bellaterra, Spain. chicagpinheiro@outlook.pt.
authors.forename: Francisca
authors.initials: F
authors.lastname: Pinheiro
authors.affiliation: Department of Biochemistry and Molecular Biology, Autonomous University of Barcelona, 08193 Bellaterra, Spain. susanna.navarro.cantero@uab.cat.|Institute of Biotechnology and Biomedicine, Autonomous University of Barcelona, 08193 Bellaterra, Spain. susanna.navarro.cantero@uab.cat.
authors.forename: Susanna
authors.initials: S
authors.lastname: Navarro
authors.affiliation: Department of Biochemistry and Molecular and Cell Biology, Institute for Biocomputation and Physics of Complex Systems (BIFI), University of Zaragoza, 50018 Zaragoza, Spain. jsancho@unizar.es.
authors.forename: Javier
authors.initials: J
authors.lastname: Sancho
authors.affiliation: Department of Biochemistry and Molecular Biology, Autonomous University of Barcelona, 08193 Bellaterra, Spain. salvador.ventura@uab.cat.|Institute of Biotechnology and Biomedicine, Autonomous University of Barcelona, 08193 Bellaterra, Spain. salvador.ventura@uab.cat.
authors.forename: Salvador
authors.initials: S
authors.lastname: Ventura
mesh_terms: D062126:Databases, Chemical; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007700:Kinetics; D010300:Parkinson Disease / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000494:pharmacology*; D051844:alpha-Synuclein / Q000037:antagonists & inhibitors*
chemical_list: D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D051844:alpha-Synuclein
keywords: Parkinson disease; amyloid; high-throughput screening; protein aggregation; α-synuclein