title: Prefibrillar transthyretin oligomers and cold stored native tetrameric transthyretin are cytotoxic in cell culture.
abstract: Recent studies suggest that soluble, oligomeric species, which are intermediates in the fibril formation process in amyloid disease, might be the key species in amyloid pathogenesis. Soluble oligomers of human wild type transthyretin (TTR) were produced to elucidate oligomer properties. Employing ThT fluorescence, time-resolved fluorescence anisotropy of pyrene-labeled TTR, chemical cross-linking, and electron microscopy we demonstrated that early formed soluble oligomers (within minutes) from A-state TTR comprised on the average 20-30 TTR monomers. When administered to neuroblastoma cells these early oligomers proved highly cytotoxic and induced apoptosis after 48 h of incubation. More mature fibrils (>24 h of fibrillation) were non-toxic. Surprisingly, we also found that native tetrameric TTR, when purified and stored under cold conditions (4 degrees C) was highly cytotoxic. The effect could be partially restored by increasing the temperature of the protein. The cytotoxic effects of native tetrameric TTR likely stems from a hitherto unexplored low temperature induced rearrangement of the tetramer conformation that possibly is related to the conformation of misfolded TTR in amyloigogenic oligomers.
authors.affiliation: IFM- Department of Chemistry, Linköping University, SE-581 83 Linköping, Sweden.
authors.forename: Karin
authors.initials: K
authors.lastname: Sörgjerd
authors.forename: Therése
authors.initials: T
authors.lastname: Klingstedt
authors.forename: Mikael
authors.initials: M
authors.lastname: Lindgren
authors.forename: Katarina
authors.initials: K
authors.lastname: Kågedal
authors.forename: Per
authors.initials: P
authors.lastname: Hammarström
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D016880:Anisotropy; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D003080:Cold Temperature; D004789:Enzyme Activation; D005453:Fluorescence; D006801:Humans; D046529:Microscopy, Electron, Transmission; D011228:Prealbumin / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology / Q000648:ultrastructure; D017510:Protein Folding
chemical_list: D000682:Amyloid; D011228:Prealbumin; D053148:Caspase 3