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Biomedical Applications of Biophysics pp 119–136Cite as

FRET and Its Biological Application as a Molecular Ruler

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Part of the book series: Handbook of Modern Biophysics ((HBBT,volume 3))

Abstract

Fluorescence resonance energy transfer (FRET) is a well-studied physical process in which two fluorescent molecules in close proximity interact with each other. During this interaction, energy is passed from one fluorophore (the donor) to the other fluorophore (the acceptor). This simple physical phenomenon can be found in many natural biological systems. For example, the luminescence of the green light-emitting Northwest Pacific jellyfish Aequorea victoriainvolves two proteins: aequorin and green fluorescent protein (GFP). The chemiluminescent aequorin emits a blue light by itself [1], but can pass its energy to GFP in the light organs of the jellyfish [2,3]. It is the fluorescence emission of GFP that yields the characteristic green light of the jellyfish.

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Authors and Affiliations

  1. Department of Physiology and Membrane Biology, School of Medicine, University of California Davis, One Shields Avenue, Davis, CA, 95616, USA

    Jie Zheng

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  1. Jie Zheng
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  1. Department of Biochemistry and Molecular Medicine, University of California Davis, One Shields Avenue, Davis, CA, 95616, USA

    Thomas Jue Ph.D.

5.1 Electronic Supplementary material

Figure 5.1.

Jablonski diagram illustrating the energy coupling process. Arrows indicate energy transitions; S 0 is the ground state, S 1 and S 2 are excited states, each with a number of vibrational energy levels; hυe represents the photon energy absorbed by the donor, hυD and hυA are photon emission from the donor and acceptor, respectively; the rate of each transition process is shown in unit of second. On the top, a diagram showing a FRET pair and light path is shown in the same colors as the energy diagram. Note, however, that energy transfer is a non-radiative process with no photon involved. Please visit http://extras.springer.com/ to view a high-resolution full-color version of this illustration. (PDF 2,775 KB)

Figure 5.2.

Number of FRET articles published each year. A PubMed search with the keyword “fluorescence resonance energy transfer” was used to construct this figure. The number of articles found in 2006 and 2007 (by mid-October) are shown. Also labeled are major events related to the fluorescent proteins. In light of the massive body of literature, examples used in this chapter will be mostly limited to ion channel studies. Please visit http://extras.springer.com/ to view a high-resolution full-color version of this illustration. (PDF 2,778 KB)

Figure 5.3.

Distance dependence of the FRET efficiency. (Left) A schematic diagram illustrating two membrane proteins each labeled with a fluorophore. When they are far apart (top), there is no FRET between the fluorophores. The fluorophores are independent of each other. When the two proteins are interacting (bottom), the fluorophores are at close proximity. Energy transfer occurs. (Right) A FRET pair with an R 0 of 50 Å is used to construct this figure. Points A, B, and C represent 50, 95, and 5% FRET efficiency, respectively. Please visit http://extras.springer.com/ to view a high-resolution full-color version of this illustration. (PDF 2,776 KB)

978-1-60327-233-9_5_MOESM4_ESM.pdf

Please visit http://extras.springer.com/ to view a high-resolution full-color version of this illustration. (PDF 2,772 KB)

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Zheng, J. (2010). FRET and Its Biological Application as a Molecular Ruler. In: Jue, T. (eds) Biomedical Applications of Biophysics. Handbook of Modern Biophysics, vol 3. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-233-9_5

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