title: Transthyretin as both a sensor and a scavenger of β-amyloid oligomers.
abstract: Transthyretin (TTR) is a homotetrameric transport protein, assembled from monomers that each contain two four-stranded β-sheets and a short α-helix and loop. In the tetramer, the "inner" β-sheet forms a hydrophobic pocket while the helix and loop are solvent-exposed. β-Amyloid (Aβ) aggregates bind to TTR, and the level of binding is significantly reduced in mutants L82A (on the loop) and L110A (on the inner β-sheet). Protection against Aβ toxicity was demonstrated for wild-type TTR but not L82A or L110A, providing a direct link between TTR-Aβ binding and TTR-mediated cytoprotection. Protection is afforded at substoichiometric (1:100) TTR:Aβ molar ratios, and the level of binding of Aβ to TTR is highest for partially aggregated materials and decreased for freshly prepared or heavily aggregated Aβ, suggesting that TTR binds selectively to soluble toxic Aβ aggregates. A novel technique, nanoparticle tracking, is used to show that TTR arrests Aβ aggregation by both preventing formation of new aggregates and inhibiting growth of existing aggregates. TTR tetramers are normally quite stable; tetrameric structure is necessary for the protein's transport functions, and mutations that decrease tetramer stability have been linked to TTR amyloid diseases. However, TTR monomers bind more Aβ than do tetramers, presumably because the hydrophobic inner sheet is solvent-exposed upon tetramer disassembly. Wild-type and L110A tetramers, but not L82A, were destabilized upon being co-incubated with Aβ, suggesting that binding of Aβ to L82 triggers tetramer dissociation. Taken together, these results suggest a novel mechanism of action for TTR: the EF helix/loop "senses" the presence of soluble toxic Aβ oligomers, triggering destabilization of TTR tetramers and exposure of the hydrophobic inner sheet, which then "scavenges" these toxic oligomers and prevents them from causing cell death.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Dennis T
authors.initials: DT
authors.lastname: Yang
authors.forename: Gururaj
authors.initials: G
authors.lastname: Joshi
authors.forename: Patricia Y
authors.initials: PY
authors.lastname: Cho
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Johnson
authors.forename: Regina M
authors.initials: RM
authors.lastname: Murphy
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism*; D001665:Binding Sites; D001704:Biopolymers / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D007700:Kinetics; D008958:Models, Molecular; D063347:Native Polyacrylamide Gel Electrophoresis; D011228:Prealbumin / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D016229:Amyloid beta-Peptides; D001704:Biopolymers; D011228:Prealbumin