title: ESI-IMS-MS: A method for rapid analysis of protein aggregation and its inhibition by small molecules.
abstract: Electrospray ionisation-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS) is a powerful method for the study of conformational changes in protein complexes, including oligomeric species populated during protein self-aggregation into amyloid fibrils. Information on the mass, stability, cross-sectional area and ligand binding capability of each transiently populated intermediate, present in the heterogeneous mixture of assembling species, can be determined individually in a single experiment in real-time. Determining the structural characterisation of oligomeric species and alterations in self-assembly pathways observed in the presence of small molecule inhibitors is of great importance, given the urgent demand for effective therapeutics. Recent studies have demonstrated the capability of ESI-IMS-MS to identify small molecule modulators of amyloid assembly and to determine the mechanism by which they interact (positive, negative, non-specific binding, or colloidal) in a high-throughput format. Here, we demonstrate these advances using self-assembly of Aβ40 as an example, and reveal two new inhibitors of Aβ40 fibrillation.
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, United Kingdom. Electronic address: bs08lw@leeds.ac.uk.
authors.forename: Lydia M
authors.initials: LM
authors.lastname: Young
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, United Kingdom. Electronic address: bsjct@leeds.ac.uk.
authors.forename: Janet C
authors.initials: JC
authors.lastname: Saunders
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, United Kingdom. Electronic address: bs08ram@leeds.ac.uk.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Mahood
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Chemistry, University of Leeds, LS2 9JT, United Kingdom. Electronic address: C.H.Revill@leeds.ac.uk.
authors.forename: Charlotte H
authors.initials: CH
authors.lastname: Revill
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Chemistry, University of Leeds, LS2 9JT, United Kingdom. Electronic address: R.Foster@leeds.ac.uk.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Foster
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, United Kingdom. Electronic address: A.E.Ashcroft@leeds.ac.uk.
authors.forename: Alison E
authors.initials: AE
authors.lastname: Ashcroft
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom; School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, United Kingdom. Electronic address: S.E.Radford@leeds.ac.uk.
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
mesh_terms: D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors* / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008958:Models, Molecular; D010446:Peptide Fragments / Q000037:antagonists & inhibitors* / Q000737:chemistry; D066329:Protein Aggregates / Q000187:drug effects*; D011485:Protein Binding; D011487:Protein Conformation; D054852:Small Molecule Libraries / Q000737:chemistry*; D012996:Solutions; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D016229:Amyloid beta-Peptides; D008024:Ligands; D010446:Peptide Fragments; D066329:Protein Aggregates; D054852:Small Molecule Libraries; D012996:Solutions; C078592:amyloid beta-protein (1-40)
keywords: Amyloid; Aβ; ESI-IMS–MS; Ligand screening; Small molecule inhibitor