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Cyclic Amplification of Prion Protein Misfolding

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Amyloid Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 849))

Abstract

Protein misfolding cyclic amplification (PMCA) is a technique that takes advantage of the nucleation-dependent prion replication process to accelerate the conversion of PrPC into PrPSc in the test tube. PMCA uses ultrasound waves to fragment the PrPSc 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 PrPSc 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.

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Acknowledgments

This work was supported in part by R01 NS049173 grant from NIH.

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Authors and Affiliations

  1. Mitchell Center for Alzheimer’s Disease and Related Brain Disorders, Department of Neurology, University of Texas Houston Medical School, Houston, TX, USA

    Marcelo A. Barria, Dennisse Gonzalez-Romero & Claudio Soto

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  1. Marcelo A. Barria
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  2. Dennisse Gonzalez-Romero
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  3. Claudio Soto
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Corresponding author

Correspondence to Claudio Soto .

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Editors and Affiliations

  1. School of Medicine, Dept. Psychiatry, New York University, First Ave. 550, New York, 10016, New York, USA

    Einar M. Sigurdsson

  2. Centro Nacional de Microbiología, Instituto de Salud Carlos III, Carretera Majadahonda-Pozuelo Km 2, Madrid, 28220, Spain

    Miguel Calero

  3. Inst. Química Física, Rocasolano, CSIC Madrid, C/ Serrano 119, Madrid, 28006, Spain

    María Gasset

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Barria, M.A., Gonzalez-Romero, D., Soto, C. (2012). Cyclic Amplification of Prion Protein Misfolding. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 849. Humana Press. https://doi.org/10.1007/978-1-61779-551-0_14

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  • DOI: https://doi.org/10.1007/978-1-61779-551-0_14

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