Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy

Author(s): Hiroshi Sakata, Motohiro Horiuchi, Izumi Takahashi, Masataka Kinjo

Journal Name: Current Pharmaceutical Biotechnology

Volume 11 , Issue 1 , 2010

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The conversion of prion protein (PrP) from the monomeric cellular isoform to the oligomeric pathological isoform is a crucial event in the pathogenesis of prion diseases. To investigate oligomer formation of PrP, enhanced green fluorescent protein (EGFP)-tagged PrP (EGFP-PrP) without the glycosylphosphatidylinositol (GPI) anchor was prepared and the oligomerization of EGFP-PrP induced by sodium dodecyl sulphate (SDS) was monitored by fluorescence correlation spectroscopy (FCS). The FCS analysis indicated that soluble oligomers were formed at 0.011% SDS. Furthermore, the combination of fluorescence cross-correlation spectroscopy (FCCS) and a panel of anti-PrP monoclonal antibodies (mAbs) revealed the conformational changes in PrP. Our studies provide a method to analyze conformational changes of proteins in solution.

Keywords: Fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy, prion protein, soluble oligomer, antibody, conformational change, epitope mapping

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Article Details

Year: 2010
Page: [87 - 95]
Pages: 9
DOI: 10.2174/138920110790725357

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