Springer Protocols: Abstract: Molecular Electron Microscopy Approaches to Elucidating the Mechanisms of Protein Fibrillogenesis

Molecular Electron Microscopy Approaches to Elucidating the Mechanisms of Protein Fibrillogenesis

By: Hilal A. Lashuel², Joseph S. Wall³

Abstract

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Electron microscopy (EM) has played a central role in our current understanding of the mechanisms underlying the pathogenesis of several amyloid diseases, including Alzheimer's disease, Parkinson's disease, and prion diseases. In this chapter, we discuss the application of various EM techniques to monitor and characterize quaternary structural changes during amyloid fibril formation in vitro and the potential of extending some of these techniques to characterizing ex vivo material. In particular, we would like to bring to the attention of the reader two very powerful molecular EM techniques that remain under utilized by researchers in the amyloid community, namely scanning transmission electron microscopy and single particle molecular averaging EM. An overview of the strength and limitations of these techniques as tools for elucidating the structural basis of amyloid fibril formation will be presented.

Affiliation(s): (2) Harvard Medical School, Center for Neurologic Diseases, Brigham and Women's Hospital and Department of Neurology, Cambridge, MA
(3) Brookhaven National Laboratory, Department of Biology, Upton, NY

Book Title: Amyloid Proteins: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 299 | Pub. Date: Dec-28-2004 | Page Range: 81-101 | DOI: 10.1385/1-59259-874-9:081

Subject: Protein Science

Key Words: Electron microscopy (EM) - scanning transmission electron microscopy (STEM) - single particle analysis - negative staining - α-synuclein, amyloid-ß (Aß) - amyloid - fibrils - protofibrils - oligomers - sedimentation velocity analytical ultracentrifugation (SVAU)

References

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