Abstract
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
Current Pharmaceutical Biotechnology
Title: Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy
Volume: 11 Issue: 1
Author(s): Hiroshi Sakata, Motohiro Horiuchi, Izumi Takahashi and Masataka Kinjo
Affiliation:
Keywords: Fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy, prion protein, soluble oligomer, antibody, conformational change, epitope mapping
Abstract: 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.
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Cite this article as:
Sakata Hiroshi, Horiuchi Motohiro, Takahashi Izumi and Kinjo Masataka, Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy, Current Pharmaceutical Biotechnology 2010; 11 (1) . https://dx.doi.org/10.2174/138920110790725357
DOI https://dx.doi.org/10.2174/138920110790725357 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
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