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Ensemble FRET Methods in Studies of Intrinsically Disordered Proteins

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Intrinsically Disordered Protein Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 895))

Abstract

The main structural characteristic of intrinsically disordered proteins (IDPs) or intrinsically disordered regions of globular proteins is that they exist as ensembles of multiple conformers which can continuously interconvert, and at times, form ensembles of a more restricted number of conformers. Characterization of the disordered state and transitions to partially or fully ordered states of such ensembles must be expressed in statistical terms, i.e., determination of probability distributions of the various conformers. This can be achieved by measurements of time-resolved dynamic non-radiative excitation energy transfer within ensembles of site-specifically labeled IDP molecules. Distributions of intramolecular segmental end-to-end distances and their fast fluctuations can be determined and fast and slow conformational transitions within selected sections of the molecule can be monitored and analyzed.

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Notes

  1. 1.

    The acronym FRET denoting fluorescence resonance energy transfer is inaccurate, because the transfer does not involve any fluorescence but non-radiative transfer of the electronic excitation energy of the donor. This mechanism does not depend on the fluorescence properties of the acceptor. The correct term should be EET representing electronic energy transfer or excitation energy transfer or RET for resonance energy transfer. However, since the term FRET has been accepted in the literature, we continue to use it here.

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  1. The Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel

    Elisha Haas

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  1. Elisha Haas
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  1. Department of Molecular Medicine, College of Medicine, University of South Florida, Bruce B. Downs Blvd 12901, Tampa, 33612, Florida, USA

    Vladimir N. Uversky

  2. Dept. Biochemistry & Molecular Biology, Center for Computational Biology &, Indiana University, West 10th Street 410, Indianapolis, 46202, Indiana, USA

    A. Keith Dunker

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Haas, E. (2012). Ensemble FRET Methods in Studies of Intrinsically Disordered Proteins. In: Uversky, V., Dunker, A. (eds) Intrinsically Disordered Protein Analysis. Methods in Molecular Biology, vol 895. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-927-3_28

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  • DOI: https://doi.org/10.1007/978-1-61779-927-3_28

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  • Publisher Name: Humana Press, Totowa, NJ

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