Conversion of Abeta42 into a folded soluble native-like protein using a semi-random library of amphipathic helices.
The amyloid cascade model hypothesizes that neurotoxic oligomers or aggregates formed by the Alzheimer amyloid peptide (Abeta) cause disease pathology in Alzheimer's disease. Attempted treatment strategies for Alzheimer's disease have involved either inhibiting Abeta oligomerization or aggregation, or dissolving existing aggregates. Blocking such downhill processes, however, has proved daunting. We have used a different approach that targets Abeta before the oligomerization cascade begins. We predicted that an amphipathic helix could convert Abeta into a native-like protein and inhibit initiation of oligomerization and aggregation. This idea was tested with a designed library and genetic screen. We exhaustively screened a library of semi-randomized amphipathic helical sequences, each expressed as a fusion protein with an Abeta42-yellow fluorescent protein sequence serving as a reporter for folding and solubilization. This yielded an amphipathic helix capable of initiating native-like folding in Abeta42 and preventing aggregation. This amphipathic helix has direct application to Alzheimer's disease therapy development.
Arslan
Pharhad Eli
PE
Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, TMDT 4-307, 101 College Street, Toronto, Ontario, Canada.
Mulligan
Vikram Khipple
VK
Ho
Sylvia
S
Chakrabartty
Avijit
A
D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry / Q000648:ultrastructure; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D017510:Protein Folding; D055503:Protein Multimerization; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility
D000682:Amyloid; D016229:Amyloid beta-Peptides; D017931:DNA Primers; D010446:Peptide Fragments; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; C075222:amyloid beta-protein (1-42); D049452:Green Fluorescent Proteins