title: Prion protein-coated magnetic beads: synthesis, characterization and development of a new ligands screening method.
abstract: Prion diseases are characterized by protein aggregation and neurodegeneration. Conversion of the native prion protein (PrP(C)) into the abnormal scrapie PrP isoform (PrP(Sc)), which undergoes aggregation and can eventually form amyloid fibrils, is a critical step leading to the characteristic path morphological hallmark of these diseases. However, the mechanism of conversion remains unclear. It is known that ligands can act as cofactors or inhibitors in the conversion mechanism of PrP(C) into PrP(Sc). Within this context, herein, we describe the immobilization of PrP(C) onto the surface of magnetic beads and the morphological characterization of PrP(C)-coated beads by fluorescence confocal microscopy. PrP(C)-coated magnetic beads were used to identify ligands from a mixture of compounds, which were monitored by UHPLC-ESI-MS/MS. This affinity-based method allowed the isolation of the anti-prion compound quinacrine, an inhibitor of PrP aggregation. The results indicate that this approach can be applied to not only "fish" for anti-prion compounds from complex matrixes, but also to screening for and identify possible cellular cofactors involved in the deflagration of prion diseases.
authors.affiliation: Universidade Federal Fluminense, Instituto de Química, Departamento de Química Orgânica, 24210-141 Niterói, RJ, Brazil; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica, 21941-902 Rio de Janeiro, RJ, Brazil. Electronic address: mcmoraes@id.uff.br.
authors.forename: Marcela Cristina
authors.initials: MC
authors.lastname: de Moraes
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Juliana Bosco
authors.initials: JB
authors.lastname: Santos
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Daniel Meira
authors.initials: DM
authors.lastname: Dos Anjos
authors.affiliation: Instituto Nacional de Ciência e Tecnologia de Biologia Estrutural e Bioimagem, 21941-902 Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Faculdade de Farmácia, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Luciana Pereira
authors.initials: LP
authors.lastname: Rangel
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica, 21941-902 Rio de Janeiro, RJ, Brazil; Instituto Nacional de Ciência e Tecnologia de Biologia Estrutural e Bioimagem, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Tuane Cristine Ramos Gonçalves
authors.initials: TC
authors.lastname: Vieira
authors.affiliation: Biomedical Research Center, National Institute on Aging, National Institutes of Health, 251 Bayview Boulevard, Baltimore, MD 21224, USA.
authors.forename: Ruin
authors.initials: R
authors.lastname: Moaddel
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica, 21941-902 Rio de Janeiro, RJ, Brazil; Instituto Nacional de Ciência e Tecnologia de Biologia Estrutural e Bioimagem, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Jerson Lima
authors.initials: JL
authors.lastname: da Silva
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D057166:High-Throughput Screening Assays / Q000295:instrumentation* / Q000379:methods*; D008024:Ligands; D060328:Magnetic Phenomena; D008856:Microscopy, Fluorescence; D018620:PrPSc Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D020033:Protein Isoforms; D011796:Quinacrine / Q000302:isolation & purification; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
chemical_list: D008024:Ligands; D018620:PrPSc Proteins; D020033:Protein Isoforms; D011796:Quinacrine
keywords: Anti-prion compounds; Ligand fishing; Magnetic beads; Prion protein; Quinacrine; Screening method