title: Screening and classifying small-molecule inhibitors of amyloid formation using ion mobility spectrometry-mass spectrometry.
abstract: The search for therapeutic agents that bind specifically to precursor protein conformations and inhibit amyloid assembly is an important challenge. Identifying such inhibitors is difficult because many protein precursors of aggregation are partially folded or intrinsically disordered, which rules out structure-based design. Furthermore, inhibitors can act by a variety of mechanisms, including specific or nonspecific binding, as well as colloidal inhibition. Here we report a high-throughput method based on ion mobility spectrometry-mass spectrometry (IMS-MS) that is capable of rapidly detecting small molecules that bind to amyloid precursors, identifying the interacting protein species and defining the mode of inhibition. Using this method we have classified a variety of small molecules that are potential inhibitors of human islet amyloid polypeptide (hIAPP) aggregation or amyloid-beta 1-40 aggregation as specific, nonspecific, colloidal or non-interacting. We also demonstrate the ability of IMS-MS to screen for inhibitory small molecules in a 96-well plate format and use this to discover a new inhibitor of hIAPP amyloid assembly.
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK.
authors.forename: Lydia M
authors.initials: LM
authors.lastname: Young
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK.
authors.forename: Janet C
authors.initials: JC
authors.lastname: Saunders
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Mahood
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Chemistry, University of Leeds, LS2 9JT, UK.
authors.forename: Charlotte H
authors.initials: CH
authors.lastname: Revill
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Chemistry, University of Leeds, LS2 9JT, UK.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Foster
authors.affiliation: Department of Chemistry, Stony Brook University, Stony Brook, NY 11794-3400, USA.
authors.forename: Ling-Hsien
authors.initials: LH
authors.lastname: Tu
authors.affiliation: Department of Chemistry, Stony Brook University, Stony Brook, NY 11794-3400, USA.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Raleigh
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK.
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK.
authors.forename: Alison E
authors.initials: AE
authors.lastname: Ashcroft
mesh_terms: D000595:Amino Acid Sequence; D000682:Amyloid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors / Q000378:metabolism; D003102:Colloids / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D058228:Islet Amyloid Polypeptide / Q000037:antagonists & inhibitors / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000378:metabolism; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000737:chemistry / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000682:Amyloid; D016229:Amyloid beta-Peptides; D003102:Colloids; D058228:Islet Amyloid Polypeptide; D010446:Peptide Fragments; D054852:Small Molecule Libraries; C078592:amyloid beta-protein (1-40)