Characterizaion of conformation-dependent prion protein epitopes

Hae-Eun Kang, Chu Chun Weng, Eri Saijo, Vicki Saylor, Jifeng Bang, Schun Kim, Laylan Ramos, Rachel Angers, Katie Langenfeld, Vadim Khaychuk, Carla Calvi, Jason Bartz, Nora Hunter, Glenn Telling

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Whereas prion replication involves structural rearrangement of cellular prion
protein (PrPC), the existence of conformational epitopes remains speculative and controversial, and PrP transformation is monitored by immunoblot detection of PrP27-30, a protease resistant counterpart of pathogenic PrPSc. We now describe the involvement of specific amino acids in conformational determinants of novel monoclonal antibodies (mAbs) raised against randomly chimeric PrP. Epitope recognition of two mAbs depended on polymorphisms controlling disease susceptibility. Detection by one, referred to as PRC5, required alanine and
asparagine at discontinuous mouse PrP residues 132 and 158, which acquire proximity when residues 126 to 218 form a structured globular domain. The discontinuous epitope of glycosylation-dependent mAb PRC7 also mapped within this domain at residues 154 and 185. In accordance with their conformational
dependence, tertiary structure perturbations compromised recognition by PRC5, PRC7, as well as previously characterized mAbs whose epitopes also reside in the globular domain, whereas conformation independent epitopes proximal or distal to this region were refractory to such destabilizing treatments. Our studies also address the paradox of how conformational epitopes remain functional following denaturing treatments, and indicate that PrPC and PrP27-30 both renature to a common structure that reconstitutes the globular domain.
Original languageEnglish
Pages (from-to)37219-37232
JournalJournal of Biological Chemistry
Volume287
Issue number44
Early online date6 Sept 2012
DOIs
Publication statusPublished - 2012

Keywords / Materials (for Non-textual outputs)

  • Prion protein
  • Amino Acid Sequence
  • ANIMALS
  • Antibodies, Monoclonal, murine-derived
  • CATTLE
  • Conserved Sequence
  • DEER
  • directed molecular evolution
  • Epitope Mapping
  • EPITOPES
  • HUMANS
  • HYBRIDOMAS
  • MICE
  • Mice, Transgenic
  • Models, Molecular
  • Molecular Sequence Data
  • Oxidation-Reduction
  • PrPC Proteins
  • Protein Binding
  • protein structure
  • recombiant fusion protein
  • saimiri
  • Sequence Deletion
  • SHEEP

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