title: Isolating toxic insulin amyloid reactive species that lack β-sheets and have wide pH stability.
abstract: Amyloid diseases, including Alzheimer's disease, are characterized by aggregation of normally functioning proteins or peptides into ordered, β-sheet rich fibrils. Most of the theories on amyloid toxicity focus on the nuclei or oligomers in the fibril formation process. The nuclei and oligomers are transient species, making their full characterization difficult. We have isolated toxic protein species that act like an oligomer and may provide the first evidence of a stable reactive species created by disaggregation of amyloid fibrils. This reactive species was isolated by dissolving amyloid fibrils at high pH and it has a mass >100 kDa and a diameter of 48 ± 15 nm. It seeds the formation of fibrils in a dose dependent manner, but using circular dichroism and deep ultraviolet resonance Raman spectroscopy, the reactive species was found to not have a β-sheet rich structure. We hypothesize that the reactive species does not decompose at high pH and maintains its structure in solution. The remaining disaggregated insulin, excluding the toxic reactive species that elongated the fibrils, returned to native structured insulin. This is the first time, to our knowledge, that a stable reactive species of an amyloid reaction has been separated and characterized by disaggregation of amyloid fibrils.
authors.affiliation: Howard P. Isermann Department of Chemical and Biological Engineering and The Center of Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA.
authors.forename: Caryn L
authors.initials: CL
authors.lastname: Heldt
authors.forename: Dmitry
authors.initials: D
authors.lastname: Kurouski
authors.forename: Mirco
authors.initials: M
authors.lastname: Sorci
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Grafeld
authors.forename: Igor K
authors.initials: IK
authors.lastname: Lednev
authors.forename: Georges
authors.initials: G
authors.lastname: Belfort
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D000818:Animals; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D016716:PC12 Cells; D055503:Protein Multimerization; D058849:Protein Refolding; D055550:Protein Stability; D017433:Protein Structure, Secondary; D051381:Rats; D012996:Solutions
chemical_list: D000682:Amyloid; D007328:Insulin; D012996:Solutions