title: The extracellular chaperone clusterin influences amyloid formation and toxicity by interacting with prefibrillar structures.
abstract: Clusterin is an extracellular chaperone present in all disease-associated extracellular amyloid deposits, but its roles in amyloid formation and protein deposition in vivo are poorly understood. The current study initially aimed to characterize the effects of clusterin on amyloid formation in vitro by a panel of eight protein substrates. Two of the substrates (Alzheimer's beta peptide and a PI3-SH3 domain) were then used in further experiments to examine the effects of clusterin on amyloid cytotoxicity and to probe the mechanism of clusterin action. We show that clusterin exerts potent effects on amyloid formation, the nature and extent of which vary greatly with the clusterin:substrate ratio, and provide evidence that these effects are exerted via interactions with prefibrillar species that share common structural features. Proamyloidogenic effects of clusterin appear to be restricted to conditions in which the substrate protein is present at a very large molar excess; under these same conditions, clusterin coincorporates with substrate protein into insoluble aggregates. However, when clusterin is present at much higher but still substoichiometric levels (e.g., a molar ratio of clusterin:substrate=1:10), it potently inhibits amyloid formation and provides substantial cytoprotection. These findings suggest that clusterin is an important element in the control of extracellular protein misfolding.
authors.affiliation: School of Biological Sciences, University of Wollongong, Wollongong, Australia.
authors.forename: Justin J
authors.initials: JJ
authors.lastname: Yerbury
authors.forename: Stephen
authors.initials: S
authors.lastname: Poon
authors.forename: Sarah
authors.initials: S
authors.lastname: Meehan
authors.forename: Brianna
authors.initials: B
authors.lastname: Thompson
authors.forename: Janet R
authors.initials: JR
authors.lastname: Kumita
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.forename: Mark R
authors.initials: MR
authors.lastname: Wilson
mesh_terms: D000682:Amyloid / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002417:Cattle; D045744:Cell Line, Tumor; D051152:Clusterin / Q000097:blood* / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma; D016874:Neurofibrillary Tangles / Q000187:drug effects / Q000473:pathology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000682:Amyloid; C495242:CLU protein, human; D051152:Clusterin; D011993:Recombinant Fusion Proteins; D019869:Phosphatidylinositol 3-Kinases