Prion Protein Protocols

ISBN-13: 9781617378317 / Angielski / Miękka / 2010 / 280 str.

Andrew F. Hill

Opis

Prion Protein Protocols brings together a collection of current protocols in the field of mammalian prion disease research. Since identification of the prion protein gene some 20 years ago, what were once thought to be rare, neurodegenerative diseases of humans and animals have become a major research area. The major interest in this field results from the epidemic of bovine spongiform encephalopathy and the subsequent human prion disease, variant Creutzfeldt Jakob disease, which still poses an unknown risk to human health. The unknowns in the field stem from the enigmatic nature of infectious prions, the infectious agent that can transmit these diseases between individuals. Although much information has been gained over the past two decades about the molecular nature of prion proteins, only very recently have several research groups begun to cast light on how to turn the normal prion protein into its aberrant, infectious form. Prion Protein Protocols brings together a collection of protocols from 13 diff- ent laboratories in five countries covering basic science and diagnostic areas of prion research. Together, the chapters provide an up-to-date collection of current methods in this unique area of neuroscience. The notes section at the end of each methods chapter provides useful insight into the experimental techniques, and they are no doubt a benefit to researchers wanting to use these technologies."

Recenzje

From the reviews:

"Interspersed are concise reviews focusing on current growth areas in the prion field. ... The protocols are clear and authoritative. For those entering the field it would be an excellent guide." (Hugh Pennington, Microbiology Today, February, 2009)

Spis treści

Chapter 1 Cell culture models to unravel prion protein function and aberrancies in prion diseases Katarina Bedecs Chapter 2 Investigation of PrPC metabolism and function in live cells: methods for studying individual cells and cell populations. Cathryn L. Haigh, David R. Brown Chapter 3 Immunodetection of PrPSc using Western and slot blotting techniques Hanna Gyllberg and Kajsa Löfgren Chapter 4 Assaying Prions in Cell Culture: The Standard Scrapie Cell Assay (SSCA) and the Scrapie Cell Assay in End Point Format (SCEPA) Sukhvir P. Mahal, Cheryl A. Demczyk, Emery W. Smith Jr., Peter-Christian Klohn and Charles Weissmann Chapter 5 Generation of cell lines propagating infectious prions and the isolation and characterization of cell derived exosomes. Laura J Vella and Andrew F Hill. Chapter 6 Neurotoxicity of prion peptides on cultured cerebellar neurons Giuseppe D. Ciccotosto, Roberto Cappai and Anthony R. White Chapter 7 Understanding the nature of prion diseases using cell-free assays. Victoria A. Lawson Chapter 8 Methods for Conversion of Prion Protein into Amyloid Fibrils. Leonid Breydo, Natallia Makarava and Ilia V. Baskakov Chapter 9 Amplification of Purified Prions In Vitro Surachai Supattapone, Nathan R. Deleault, and Judy R. Rees Chapter 10 Expression and Purification of Full-Length Recombinant PrP of High Purity Natallia Makarava and Ilia V. Baskakov Chapter 11 Analysis of PrP conformation using circular dichroism Sen Han, and Andrew F. Hill Chapter 12 Effect of copper on the de novo generation of prion protein expressed in Pichia pastoris CarinaTreiber Chapter 13 Biophysical Investigations of the prion protein using Electron Paramagnetic Resonance Simon C. Drew and Kevin J. Barnham Chapter 14 Molecular diagnosis of human prion disease Jonathan D.F. Wadsworth, Caroline Powell, Jonathan A. Beck, Susan Joiner, Jacqueline M. Linehan, Sebastian Brandner, Simon Mead and John Collinge Chapter 15 Analysis of endogenous PrPC processing in neuronal and non-neuronal cell lines. Victoria Lewis and Steven J Collins Chapter 16 Molecular typing of PrPres in human sporadic CJD brain tissue. Victoria Lewis, Genevieve M Klug, Andrew F Hill and Steven J Collins Chapter 17 Transgenic mouse models of prion diseases Glenn Telling Chapter 18 Quantitative bioassay of surface bound prion infectivity. Victoria A. Lawson

Blurb

While much information has been gained over the last two decades regarding the molecular nature of prion proteins, only recently have several research groups begun to cast light on turning the normal prion protein into its aberrant, infectious form. In Prion Protein Protocols, an international team of experts provide an up-to-date collection of current methods in this unique area of neuroscience.  The chapters contained in this volume, both protocols and useful review chapters, feature topics such as the cell biology of prions, techniques and approaches to studying prion infection in cultured cells, how these systems can be used as a rapid bioassay, and prion protein misfolding. As a volume in the highly successful Methods in Molecular Biology™ series, the chapters include readily reproducible, step-by-step laboratory protocols, lists of the necessary materials and reagents, and the Notes section, which provides tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Prion Protein Protocols is an ideal collection for researchers investigating the growing field of mammalian prion disease.