Cyclic amplification of prion protein misfolding

Marcelo A Barria, Dennisse Gonzalez-Romero, Claudio Soto

Research output: Contribution to journalArticlepeer-review

Abstract

Protein misfolding cyclic amplification (PMCA) is a technique that takes advantage of the nucleation-dependent prion replication process to accelerate the conversion of PrP(C) into PrP(Sc) in the test tube. PMCA uses ultrasound waves to fragment the PrP(Sc) polymers, increasing the amount of seeds present in the infected sample without affecting their ability to act as conversion nuclei. Over the past 5 years, PMCA has become an invaluable technique to study diverse aspects of prions. The PMCA technology has been used by several groups to understand the molecular mechanism of prion replication, the cellular factors involved in prion propagation, the intriguing phenomena of prion strains and species barriers, to detect PrP(Sc) in tissues and biological fluids, and to screen for inhibitors against prion replication. In this chapter, we describe a detailed protocol of the PMCA technique, highlighting some of the important technical aspects to obtain a successful and reproducible application of the technology.

Original languageEnglish
Pages (from-to)199-212
Number of pages14
JournalMethods in Molecular Biology
Volume849
DOIs
Publication statusPublished - 2012

Keywords

  • Animals
  • Brain
  • Cattle
  • Cricetinae
  • Humans
  • Mice
  • Prion Diseases
  • Prions
  • Protein Folding
  • Sonication

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