Protein Forster resonance energy transfer

Liao F, Xie Y, Yang X et al (2009) Homogeneous noncompetitive assay of protein via Forster-resonance-energy-transfer with tryptophan residue(s) as intrinsic donor(s) and fluorescent ligand as acceptor. Biosens Bioelectron 25 112-117... 

Day RN, Booker CF, Periasamy A (2008) Characterization of an improved donor fluorescent protein for Forster resonance energy transfer microscopy. J Biomed Opt 13 031203. doi 10.1117/1111.2939094... 

The lifetime of the excited state of fluorophores may be altered by physical and biochemical properties of its environment. Fluorescence lifetime imaging microscopy (FLIM) is thus a powerful analytical tool for the quantitative mapping of fluorescent molecules that reports, for instance, on local ion concentration, pH, and viscosity, the fluorescence lifetime of a donor fluorophore, Forster resonance energy transfer can be also imaged by FLIM. This provides a robust method for mapping protein-protein interactions and for probing the complexity of molecular interaction networks. 

Ganesan, S., Ameer-Beg, S. M., Ng, T. T., Vojnovic, B. and Wouters, F. S. (2006). A dark yellow fluorescent protein (YFP)-based Resonance Energy-Accepting Chromoprotein (REACh) for Forster resonance energy transfer with GFP. Proc. Natl. Acad. Sci. USA 103, 4089-94. 

Fluorescence or Forster resonance energy transfer (FRET) is widely accepted as being one of the most useful methods to observe biochemical events in vitro and in living cells. Generally, there are two forms of FRET sensors those based on a pair of genetically encoded fluorophores, usually employing fluorescent proteins from jellyfish or corals, or those based on small molecules that make use of small organic fluorophores. 

Key words Forster resonance energy transfer. Image cytometry. Protein interactions, Pertuziunab,... 

More sophisticated designs involved semiconductor quantum dots with fluorescent protein receptors immobilized on the surface [146], The binding site of the protein receptor is occupied with an efficient fluorophore. On excitation a series of FRET (Forster resonant energy transfer) processes takes place excitation energy is transferred from the core of the quantum dot to the fluorescent protein and subsequently to the fluorophore. On substrate binding only one FRET step takes place and luminescence of the receptor is observed [146], In the simplest sensor architecture the protein contains bound quencher. Upon interaction with analyte the quencher is liberated and luminescence of the quantum dot is observed (Figure 16.25c). 

Forster Resonance Energy Transfer (FRET) for Proteins... 

Merzlyakov M, Li E, Casas R, Hristova K. Spectral Forster resonance energy transfer detection of protein interactions in surface-supported bilayers. Langmuir 2006 22 6986-6992. 

S.Y. Breusegem, In vivo investigation of protein interactions in C. Elegans by Forster resonance energy transfer microscopy, University of Illinois at Urbana-Champaign (2002)... 

While the complementary technique Forster resonant energy transfer (FRET), which is widely used for studying distances in proteins requires two different, relatively large chromophores, which must be chosen according to the expected distance, EPR distance measurements can be performed using two identical much smaller nitroxide labels and are precise over a broad range of distances [51,65,66]. 

Key words Confocal fluorescence microscopy, Forster resonance energy transfer, Total internal reflection fluorescence microscopy, Single-molecule imaging, GPCR, Heterotrimeric G-proteins, Spatiotemporal dynamics... 

Second, we present a Forster resonance energy transfer (FRET) imaging method, which has been used to monitor GPGR-mediated dissociation (activation) and reassociation (deactivation) of heterotrimeric G-protein in single live cells (10, 11). Protein/protein interactions cannot be measured by colocalization of the proteins because the limit of resolution of the light microscope using standard techniques is on the order of hundreds of nanometers, and one cannot be certain that two proteins of interest physically interact even... 

In fact, FRET (Forster resonant energy transfer) analysis has the same basis as the energy transfer described in the above section. It is used either in simple bioanalyses or to detect protein interactions and DNA hybridization. Its principle is shown on Fig. 14 in the case of a homogeneous immunoassay [43]. 

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