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Lookup NU author(s): Professor Tiago Outeiro
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The main feature of protein misfolding disorders is the presence of protein deposits in cells and tissues as a consequence of aberrant protein-protein interactions. Neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease belong to this class of disorders. The deposition of large amyloid deposits is a dynamic process that starts with the generation of small, soluble species known as dimers and oligomers. The identification of physical, chemical and genetic conditions that promote - or inhibit - the pathologic deposition of proteins is essential for understanding and preventing neurodegeneration. In this chapter, we explain the history, features, design, development and optimization of bimolecular fluorescence complementation (BiFC) assays for the study of protein oligomerization, aggregation and toxicity in neurodegenerative disorders. In BiFC assays protein chimeras containing the protein of interest fused to non-fluorescent halves of a fluorescent reporter protein are generated. When the proteins of interest interact, the halves of the fluorophore become close enough to reconstitute the fluorescence, which is therefore proportional to the production of dimers and oligomers of the proteins of interest. The fact that BiFC assays allow the direct visualization and measurement of the first steps of aggregation makes them a unique tool in the field of protein misfolding disorders. Additionally, BiFC assays require only basic cell and molecular biology materials and equipment, have a wide array of possible experimental settings/fluorophores, allow studies in living cells and organisms, and are especially suitable for large drug and genetic screenings. © 2014 Elsevier Inc. All rights reserved..
Author(s): Herrera F, Goncalves S, Branco dos Santos J, Fleming Outeiro T
Publication type: Book Chapter
Publication status: Published
Book Title: Bio-nanoimaging: Protein Misfolding and Aggregation
Online publication date: 22/11/2013
Acceptance date: 01/01/1900
Publisher: Elsevier Inc.
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