Conformational detection of prion protein with biarsenical labeling and FIAsH fluorescence

Coleman, Bradley M., Nisbet, Rebecca M., Han, Sen, Cappai, Roberto, Hatters, Danny M. and Hill, Andrew F. (2009) Conformational detection of prion protein with biarsenical labeling and FIAsH fluorescence. Biochemical and Biophysical Research Communications, 380 3: 564-568. doi:10.1016/j.bbrc.2009.01.120

Author Coleman, Bradley M.
Nisbet, Rebecca M.
Han, Sen
Cappai, Roberto
Hatters, Danny M.
Hill, Andrew F.
Title Conformational detection of prion protein with biarsenical labeling and FIAsH fluorescence
Journal name Biochemical and Biophysical Research Communications   Check publisher's open access policy
ISSN 1090-2104
Publication date 2009-03-13
Sub-type Article (original research)
DOI 10.1016/j.bbrc.2009.01.120
Open Access Status
Volume 380
Issue 3
Start page 564
End page 568
Total pages 5
Place of publication Philadelphia, United States
Publisher Elsevier
Language eng
Formatted abstract
Prion diseases are associated with the misfolding of the host-encoded cellular prion protein (PrPC) into a disease associated form (PrPSc). Recombinant PrP can be refolded into either an α-helical rich conformation (α-PrP) resembling PrPC or a β-sheet rich, protease resistant form similar to PrPSc. Here, we generated tetracysteine tagged recombinant PrP, folded this into α- or β-PrP and determined the levels of FlAsH fluorescence. Insertion of the tetracysteine tag at three different sites within the 91–111 epitope readily distinguished β-PrP from α-PrP upon FlAsH labeling. Labelling of tetracysteine tagged PrP in the α-helical form showed minimal fluorescence, whereas labeling of tagged PrP in the β-sheet form showed high fluorescence indicating that this region is exposed upon conversion. This highlights a region of PrP that can be implicated in the development of diagnostics and is a novel, protease free mechanism for distinguishing PrPSc from PrPC. This technique may also be applied to any protein that undergoes conformational change and/or misfolding such as those involved in other neurodegenerative disorders including Alzheimer’s, Huntington’s and Parkinson’s diseases.
Keyword Prion
Protein folding
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Brain Institute Publications
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Citation counts: TR Web of Science Citation Count  Cited 23 times in Thomson Reuters Web of Science Article | Citations
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Created: Thu, 06 Mar 2014, 13:58:48 EST by Rebecca Nisbet on behalf of Clem Jones Centre for Ageing Dementia Research