Intra- and Interspecies Interactions between Prion Proteins and Effects of Mutations and Polymorphisms
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Abstract
Recently, crystallization of the prion protein in a dimeric form was reported. Here we show that native soluble homogenous FLAG-tagged prion proteins from hamster, man and cattle expressed in the baculovirus system are predominantly dimeric. The PrP/PrP interaction was confirmed in Semliki Forest virus-RNA transfected BHK cells co-expressing FLAG and oligohistidine-tagged human PrP. The yeast two-hybrid system identified the octarepeat region and the C-terminal structured domain (aa90- aa230) of PrP as PrP/PrP interaction domains. Additional octarepeats identified in patients suffering from fCJD reduced (wtPrP versus PrP+9OR) and completely abolished (PrP+9OR versus PrP+9OR) the PrP/PrP interaction in the yeast two-hybrid system. In contrast, the Met/Val polymorphism (aa129), the GSS mutation Pro102Leu and the FFI mutation Asp178Asn did not affect PrP/PrP interactions. Proof of interactions between human or sheep and bovine PrP, and sheep and human PrP, as well as lack of interactions between human or bovine PrP and hamster PrP suggest that interspecies PrP interaction studies in the yeast two-hybrid system may serve as a rapid pre-assay to investigate species barriers in prion diseases.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- The Circadian Clock of the Unicellular Eukaryotic Model Organism Chlamydomonas reinhardtii
- Cardiovascular Control by the Suprachiasmatic Nucleus: Neural and Neuroendocrine Mechanisms in Human and Rat
- Synchronization of the Molecular Clockwork by Light- and Food-Related Cues in Mammals
- Identification of Rhythmic Subsystems in the Circadian Cycle of Crassulacean Acid Metabolism under Thermoperiodic Perturbations
- Age-Dependent Effects of Conditioning on Cholinergic and Vasopressin Systems in the Rat Suprachiasmatic Nucleus
- The Magnetofection Method: Using Magnetic Force to Enhance Gene Delivery
- The Protease Inhibitor Bikunin, a Novel Anti-Metastatic Agent
- Transcriptional Regulation of the Urokinase Receptor (u-PAR) – A Central Molecule of Invasion and Metastasis
- Identification and Characterization of KAT, a Novel Gene Preferentially Expressed in Several Human Cancer Cell Lines
- Trichostatin A and Structurally Related Histone Deacetylase Inhibitors Induce 5-Lipoxygenase Promoter Activity
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- Fabin, a Novel Calcyon-Like and Glucanase-Like Protein with Mitogenic, Antifungal and Translation-Inhibitory Activities from Broad Beans
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