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Biophys J. 2015 Aug 18;109(4):793-805. doi: 10.1016/j.bpj.2015.06.065.

Prion Protein-Antibody Complexes Characterized by Chromatography-Coupled Small-Angle X-Ray Scattering.

Author information

  • 1Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California.
  • 2Department of Bioengineering and Therapeutic Sciences and Department of Pharmaceutical Chemistry and California Institute for Quantitative Biosciences (QB3), University of California San Francisco, San Francisco, California.
  • 3Institute for Neurodegenerative Diseases, University of California San Francisco, San Francisco, California; Department of Neurology, University of California San Francisco, San Francisco, California.
  • 4Institute for Neurodegenerative Diseases, University of California San Francisco, San Francisco, California.
  • 5Institute for Neurodegenerative Diseases, University of California San Francisco, San Francisco, California; Department of Neurology, University of California San Francisco, San Francisco, California. Electronic address: stanley@ind.ucsf.edu.
  • 6Department of Bioengineering and Therapeutic Sciences and Department of Pharmaceutical Chemistry and California Institute for Quantitative Biosciences (QB3), University of California San Francisco, San Francisco, California. Electronic address: sali@salilab.org.

Abstract

Aberrant self-assembly, induced by structural misfolding of the prion proteins, leads to a number of neurodegenerative disorders. In particular, misfolding of the mostly α-helical cellular prion protein (PrP(C)) into a β-sheet-rich disease-causing isoform (PrP(Sc)) is the key molecular event in the formation of PrP(Sc) aggregates. The molecular mechanisms underlying the PrP(C)-to-PrP(Sc) conversion and subsequent aggregation remain to be elucidated. However, in persistently prion-infected cell-culture models, it was shown that treatment with monoclonal antibodies against defined regions of the prion protein (PrP) led to the clearing of PrP(Sc) in cultured cells. To gain more insight into this process, we characterized PrP-antibody complexes in solution using a fast protein liquid chromatography coupled with small-angle x-ray scattering (FPLC-SAXS) procedure. High-quality SAXS data were collected for full-length recombinant mouse PrP [denoted recPrP(23-230)] and N-terminally truncated recPrP(89-230), as well as their complexes with each of two Fab fragments (HuM-P and HuM-R1), which recognize N- and C-terminal epitopes of PrP, respectively. In-line measurements by fast protein liquid chromatography coupled with SAXS minimized data artifacts caused by a non-monodispersed sample, allowing structural analysis of PrP alone and in complex with Fab antibodies. The resulting structural models suggest two mechanisms for how these Fabs may prevent the conversion of PrP(C) into PrP(Sc).

PMID:
26287631
PMCID:
PMC4547163
DOI:
10.1016/j.bpj.2015.06.065
[PubMed - indexed for MEDLINE]
Free PMC Article
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