Non-glycosylphosphatidylinositol (GPI)-anchored recombinant prion protein with dominant-negative mutation inhibits PrPSc replication in vitro

Hitaru Kishida, Yuji Sakasegawa, Kota Watanabe, Yoshio Yamakawa, Masahiro Nishijima, Yoshiyuki Kuroiwa, Naomi S. Hachiya, Kiyotoshi Kaneko

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

Dominant-negative mouse prion protein (PrP) with a lysine mutation at codon 218 (Q218K) is known to inhibit prion replication. In order to gain further mechanistic insight into such dominant negative inhibition, non- glycosylphosphatidylinositol (GPI)-anchored recombinant PrP with Q218K (rPrP-Q218K) was investigated. When applied into scrapie-infected mouse neuroblastoma (ScN2a) cells, rPrP-Q218K but not wild-type rPrP (rPrP-WT) exclusively inhibited abnormal protease-resistant pathogenic isoform (PrP Sc) replication without reducing the viability of the cells. It was even more efficient than quinacrine, which has already been prescribed for sporadic Creutzfeldt-Jakob disease (CJD) patients; 50% effective concentration (EC50) = 0.20 μM, 99% effective concentration (EC99) = 0.86 μM vs. EC50 = 0.45 μM, EC99 = 1.5 μM. Besides, no apparent cell damage was observed at the concentration of up to 4.3 μM (100 μg/ ml). In combination treatment with 0.43 μM (10 μg/ml) of rPrP-Q218K, EC99 of quinacrine was decreased from 1.5 μM to 0.5 μM, and the cell viability was recovered from 50% to over 90% as inversely proportional to the concentration of quinacrine. Such combination could alleviate the side effects of quinacrine by reducing its effective concentration without changing or even acceleration the inhibition efficacy. Since homogeneous, high-quality rPrPs could be easily prepared from Escherichia coli in large quantities, rPrP-Q218K is a good candidate for a prion replication antagonist.

Original languageEnglish
Pages (from-to)14-20
Number of pages7
JournalAmyloid
Volume11
Issue number1
DOIs
Publication statusPublished - 2004 Mar

Keywords

  • Creutzfeldt-Jakob disease (CJD)
  • Dominant negatives
  • Glycosylphosphatidylinositol (GPI)-anchor
  • Lipid rafts
  • Q218K
  • Quinacrine
  • Recombinant prion protein (rPrP)

ASJC Scopus subject areas

  • Internal Medicine

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