title: Intracellular selection of peptide inhibitors that target disulphide-bridged Aβ42 oligomers.
abstract: The β-amyloid (Aβ) peptide aggregates into a number of soluble and insoluble forms, with soluble oligomers thought to be the primary factor implicated in Alzheimer's disease pathology. As a result, a wide range of potential aggregation inhibitors have been developed. However, in addition to problems with solubility and protease susceptibility, many have inadvertently raised the concentration of these soluble neurotoxic species. Sandberg et al. previously reported a β-hairpin stabilized variant of Aβ42 that results from an intramolecular disulphide bridge (A21C/A31C; Aβ42cc), which generates highly toxic oligomeric species incapable of converting into mature fibrils. Using an intracellular protein-fragment complementation (PCA) approach, we have screened peptide libraries using E. coli that harbor an oxidizing environment to permit cytoplasmic disulphide bond formation. Peptides designed to target either the first or second β-strand have been demonstrated to bind to Aβ42cc, lower amyloid cytotoxicity, and confer bacterial cell survival. Peptides have consequently been tested using wild-type Aβ42 via ThT binding assays, circular dichroism, MTT cytotoxicity assays, fluorescence microscopy, and atomic force microscopy. Results demonstrate that amyloid-PCA selected peptides function by both removing amyloid oligomers as well as inhibiting their formation. These data further support the use of semirational design combined with intracellular PCA methodology to develop Aβ antagonists as candidates for modification into drugs capable of slowing or even preventing the onset of AD.
authors.affiliation: School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, United Kingdom.
authors.forename: Nicola
authors.initials: N
authors.lastname: Acerra
authors.forename: Neil M
authors.initials: NM
authors.lastname: Kad
authors.forename: Harish
authors.initials: H
authors.lastname: Cheruvara
authors.forename: Jody M
authors.initials: JM
authors.lastname: Mason
mesh_terms: D000544:Alzheimer Disease / Q000188:drug therapy / Q000517:prevention & control; D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D004220:Disulfides / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology*; D015202:Protein Engineering
chemical_list: D016229:Amyloid beta-Peptides; D004220:Disulfides; D019151:Peptide Library; D010455:Peptides
keywords: Alzheimer's disease; amyloid; library screening; protein misfolding; protein-fragment complementation assay; protein-protein interactions