title: Atomic force microscopy based nanoassay: a new method to study α-Synuclein-dopamine bioaffinity interactions.
abstract: Intrinsically Disordered Proteins (IDPs) are characterized by the lack of well-defined 3-D structure and show high conformational plasticity. For this reason, they are a strong challenge for the traditional characterization of structure, supramolecular assembly and biorecognition phenomena. We show here how the fine tuning of protein orientation on a surface turns useful in the reliable testing of biorecognition interactions of IDPs, in particular α-Synuclein. We exploited atomic force microscopy (AFM) for the selective, nanoscale confinement of α-Synuclein on gold to study the early stages of α-Synuclein aggregation and the effect of small molecules, like dopamine, on the aggregation process. Capitalizing on the high sensitivity of AFM topographic height measurements we determined, for the first time in the literature, the dissociation constant of dopamine-α-Synuclein adducts.
authors.affiliation: 1] NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy [2] Life Science Department, University of Trieste, via Giorgieri 1, I-34127 Trieste, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Corvaglia
authors.affiliation: 1] NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy [2] Department of Biological and Medical Science, University of Udine, Ospedale della Misericordia, Piazzale Santa Maria della Misericordia, 15 -33100 Udine, Italy [3].
authors.forename: Barbara
authors.initials: B
authors.lastname: Sanavio
authors.affiliation: Department of Biological and Medical Science, University of Udine, Ospedale della Misericordia, Piazzale Santa Maria della Misericordia, 15 -33100 Udine, Italy.
authors.forename: Rolando P
authors.initials: RP
authors.lastname: Hong Enriquez
authors.affiliation: 1] Center for Bio-Molecular Nanotechnologies@UniLe, Istituto Italiano di Tecnologia, Via Barsanti-73010 Arnesano, Lecce, Italy [2].
authors.forename: Barbara
authors.initials: B
authors.lastname: Sorce
authors.affiliation: NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bosco
authors.affiliation: 1] NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy [2] Life Science Department, University of Trieste, via Giorgieri 1, I-34127 Trieste, Italy.
authors.forename: Denis
authors.initials: D
authors.lastname: Scaini
authors.affiliation: Center for Bio-Molecular Nanotechnologies@UniLe, Istituto Italiano di Tecnologia, Via Barsanti-73010 Arnesano, Lecce, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Sabella
authors.affiliation: Center for Bio-Molecular Nanotechnologies@UniLe, Istituto Italiano di Tecnologia, Via Barsanti-73010 Arnesano, Lecce, Italy.
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Pompa
authors.affiliation: 1] NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy [2] Department of Biological and Medical Science, University of Udine, Ospedale della Misericordia, Piazzale Santa Maria della Misericordia, 15 -33100 Udine, Italy.
authors.forename: Giacinto
authors.initials: G
authors.lastname: Scoles
authors.affiliation: NanoInnovation Laboratory, Elettra Sincrotrone Trieste S.C.p.A., S.S.14 Km 163.5, 34149 Basovizza, Trieste, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Casalis
mesh_terms: D000327:Adsorption; D001665:Binding Sites; D004298:Dopamine / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D012680:Sensitivity and Specificity; D051844:alpha-Synuclein / Q000737:chemistry*
chemical_list: D051844:alpha-Synuclein; D006046:Gold; D004298:Dopamine