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Tetracysteine-tagged prion protein allows discrimination between the native and converted formsGašperšič, Jernej ...The conformational conversion of prion protein (PrP) from a native conformation to the amyloid form is a hallmark of transmissible spongiform encephalopathies. Conversion is usually monitored by ... fluorescent dyes, which bind generic amyloids and are less suited for living cell imaging. We report a new method for the synthesis of membrane-permeable and membrane-impermeable biarsenical reagents, which are then used to monitor murine PrP (mPrP) misfolding. We introduced tetracysteine (TC) tags into three different positions of mPrP, which folded into a native-like structure. Whereas mPrPs with a TC tag inserted at the N-terminus or C-terminus supported fibril formation, insertion into the helix 2-helix 3 loop inhibited conversion. We devised a quantitative protease-free method to determine the fraction of converted PrP, based on the ability of the fluorescein arsenical helix binder reagent to differentiate between the monomeric and fibrilized form of TC-tagged PrP, and showed that TC-tagged mPrP could be detected on transfectedcells, thereby expanding the potential use of this method for the detection and study of conformational diseases.Source: FEBS journal. - ISSN 1742-464X (Vol. 277, no. 9, 2010, str. 2038-2050)Type of material - article, component partPublish date - 2010Language - englishCOBISS.SI-ID - 4377626
Author
Gašperšič, Jernej |
Hafner Bratkovič, Iva, 1978- |
Stephan, Michel |
Veranič, Peter |
Benčina, Mojca, 1968- |
Vorberg, Ina |
Jerala, Roman
Topics
biarsenical |
conversion |
fibril |
prion |
tetracysteine
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Gašperšič, Jernej | 28558 |
Hafner Bratkovič, Iva, 1978- | 23563 |
Stephan, Michel | 25361 |
Veranič, Peter | 08279 |
Benčina, Mojca, 1968- | 14360 |
Vorberg, Ina | ![]() |
Jerala, Roman | 06628 |
Source: Personal bibliographies
and: SICRIS
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