title: Small molecule screening in context: lipid-catalyzed amyloid formation.
abstract: Islet Amyloid Polypeptide (IAPP) is a 37-residue hormone cosecreted with insulin by the β-cells of the pancreas. Amyloid fiber aggregation of IAPP has been correlated with the dysfunction and death of these cells in type II diabetics. The likely mechanisms by which IAPP gains toxic function include energy independent cell membrane penetration and induction of membrane depolarization. These processes have been correlated with solution biophysical observations of lipid bilayer catalyzed acceleration of amyloid formation. Although the relationship between amyloid formation and toxicity is poorly understood, the fact that conditions promoting one also favor the other suggests related membrane active structural states. Here, a novel high throughput screening protocol is described that capitalizes on this correlation to identify compounds that target membrane active species. Applied to a small library of 960 known bioactive compounds, we are able to report identification of 37 compounds of which 36 were not previously reported as active toward IAPP fiber formation. Several compounds tested in secondary cell viability assays also demonstrate cytoprotective effects. It is a general observation that peptide induced toxicity in several amyloid diseases (such as Alzhiemer's and Parkinson's) involves a membrane bound, preamyloid oligomeric species. Our data here suggest that a screening protocol based on lipid-catalyzed assembly will find mechanistically informative small molecule hits in this subclass of amyloid diseases.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, 06520-8114.
authors.forename: James A
authors.initials: JA
authors.lastname: Hebda
authors.forename: Mazin
authors.initials: M
authors.lastname: Magzoub
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Miranker
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006801:Humans; D058228:Islet Amyloid Polypeptide / Q000737:chemistry* / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D008622:Merbromin / Q000494:pharmacology*; D051381:Rats; D054852:Small Molecule Libraries
chemical_list: D000682:Amyloid; D058228:Islet Amyloid Polypeptide; D008055:Lipids; D054852:Small Molecule Libraries; D008622:Merbromin
keywords: HTF; IAPP; Type 2 diabetes; amylin; amyloid; high throughput screen; lipids; small molecules; toxicity