Recombinant human prion protein fragment 90-231, a useful model to study prion neurotoxicity

• Published in: Omics (Larchmont, N.Y.) Vol. 16; no. 1-2; p. 50
• Main Authors: Corsaro, Alessandro, Thellung, Stefano, Villa, Valentina, Nizzari, Mario, Aceto, Antonio, Florio, Tullio
• Format: Journal Article
• Language: English
• Published: United States 01.02.2012
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Cell Line - drug effects; Enzyme Inhibitors - pharmacology; Extracellular Signal-Regulated MAP Kinases - metabolism; Humans; Minocycline - pharmacology; p38 Mitogen-Activated Protein Kinases - metabolism; Peptide Fragments - chemistry; Peptide Fragments - genetics; Peptide Fragments - toxicity; Prion Diseases - etiology; Prion Diseases - pathology; Prions - chemistry; Prions - genetics; Prions - toxicity; Protein Conformation; Protein Denaturation; Protein Isoforms - chemistry; Protein Isoforms - genetics; Protein Isoforms - toxicity; Quinacrine - pharmacology; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - toxicity
• ISSN: 1557-8100
• DOI: 10.1089/omi.2011.0038

Transmissible spongiform encephalopathies (TSE), or prion diseases, are a group of fatal neurodegenerative disorders of animals and humans. Human diseases include Creutzfeldt-Jakob (CJD) and Gerstmann-Straussler-Scheinker (GSSD) diseases, fatal familial insomnia, and Kuru. Human and animal TSEs share a common histopathology with a pathognomonic triad: spongiform vacuolation of the grey matter, neuronal death, glial proliferation, and, more inconstantly, amyloid deposition. According to the "protein only" hypothesis, TSEs are caused by a unique post-translational conversion of normal, host-encoded, protease-sensitive prion protein (PrP(sen) or PrP(C)) to an abnormal disease-associated isoform (PrP(res) or PrP(Sc)). To investigate the molecular mechanism of neurotoxicity induced by PrP(Sc) we developed a protocol to obtain millimolar amounts of soluble recombinant polypeptide encompassing the amino acid sequence 90-231 of human PrP (hPrP90-231). This protein corresponds to the protease-resistant prion protein fragment that originates after amino-terminal truncation. Importantly, hPrP90-231 has a flexible backbone that, similar to PrP(C), can undergo to structural rearrangement. This peptide, structurally resembling PrP(C), can be converted in a PrP(Sc)-like conformation, and thus represents a valuable model to study prion neurotoxicity. In this article we summarized our experimental evidence on the molecular and structural mechanisms responsible of hPrP90-231 neurotoxicity on neuroectodermal cell line SHSY5Y and the effects of some PrP pathogen mutations identified in familial TSE.