Fluorescence energy transfer applications

Jovin, T. and Arndt-Jovin, D. (1989). FRET microscopy Digital imaging of fluorescence resonance energy transfer. Application in cell biology. In Cell Structure and Function by Microspectrofluorometry (Kohen, E., JG, H. and Ploem, J., eds.). Academic Press, London, pp. 99-117. 

The fluorescence energy transfer process has been widely used to determine the distance between fluorophores, the surface density of fluorophores in the lipid bilayer, and the orientation of membrane protein or protein segments, often with reference to the membrane surface and protein-protein interactions. Membranes are intrinsically dynamic in nature, so that so far the major applications have been the determination of fixed distances between molecules of interest in the membrane. 

E. A. Haigh, K. R. Thulborn, and W. H. Sawyer, Comparison of fluorescence energy transfer and quenching methods to establish the position and orientation of components within the transverse plane of the lipid bilayer. Application to the gramicidin A-bilayer interaction, Biochemistry 18, 3525-3532 (1979). 

A very interesting application of local spectroscopy on the singlc-mol-ecule level was reported by Ha et al. [99], They investigated the fluorescence energy transfer between a pair of closely spaced dye molecules, one being... 

Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis Anal. Biochem. 231 131 0... 

The application of total internal reflection fluorescence spectroscopy (TIRF) by this laboratory to the study of protein adsorption at solid-liquid interfaces is reviewed. TIRF has been used to determine adsorption isotherms and adsorption rates from single-and multi-component protein solutions. Initial adsorption rates of BSA can be explained qualitatively by the properties of the adsorbing surface. Most recently, a TIRF study using monoclonal antibodies to probe the conformation of adsorbed sperm whale myoglobin (Mb) elucidated two aspects of the Mb adsorption process 1) Mb adsorbs in a non-random manner. 2) Conformational changes of adsorbed Mb, if they occur, are minor and confined to local regions of the molecule. Fluorescence energy transfer and proteolytic enzyme techniques, when coupled with TIRF, can characterize, respectively, the conformation and orientation of adsorbed Mb. 

It is the aim of this chapter to review the most widely used techniques for time-resolved fluorescence in the life sciences, to point out possible future developments, and to give examples of typical and important applications. The reader less familiar with fluorescence techniques is referred to the chapters on fluorescence energy transfer by P. R. Selvin (this volume... 

Ju, J., Kheterpal, L, Scherer, J.R., Ruan, C., Fuller, C.W., Glazer, A.N., and Mathies, R.A., Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis, AnaZ. Biochem., 231,131,1995. 

Rogers JMG, Lippert LG, Gai F (2010) Non-natural amino acid fluorophores for one- and two-step fluorescence resonance energy transfer applications. Anal. Biochem. 399 182-9... 

On-bead sensors have also been developed on the basis of the fluorescence energy transfer (FRET) mechanism (Figure 14.2b). FRET signaling is less stringent than PET in terms of binding domain structure but requires large changes in the distance between donor and acceptor, which results in their main application within peptide or nucleic acid structures. On-bead FRET sensors constructed by split-and-mix techniques were initially... 

Oswald B, Gruber M, Bohmer M, Lehmann F, Probst M, Wolfbeis OS (2001) Novel diode laser-compatible fluorophores and their application to single molecule detection, protein labeling and fluorescence resonance energy transfer immunoassay. Photochem Photobiol 74 237-245... 

Another method to detect energy transfer directly is to measure the concentration or amount of acceptor that has undergone an excited state reaction by means other than detecting its fluorescence. For instance, by chemical analysis or chromatographic analysis of the product of a reaction involving excited A [117, 118]. An early application of this determined the photolyzed A molecules by absorption spectroscopic analysis. [119-121], This can be a powerful method, because it does not depend on expensive instrumentation however, it lacks real-time observation, and requires subsequent manipulation. For this reason, fluorescence is the usual method of detection of the sensitized excitation of the acceptor. If it is possible to excite the donor without exciting the acceptor, then the rate of photolysis of the acceptor (which is an excited state reaction) can be used to calculate the FRET efficiency [122],... 

Szolldsi, J., Damjanovich, S. and Matyus, L. (1998). Application of fluorescence resonance energy transfer in the clinical laboratory Routine and research. Cytometry 34, 159-79. 

Didenko, V. V. (2001). DNA probes using fluorescence resonance energy transfer (FRET) Designs and applications. Biotechniques 31,1106-1116. 

Blagoi, G., Rosenzweig, N. and Rosenzweig, Z. (2005). Design, synthesis, and application of particle-based fluorescence resonance energy transfer sensors for carbohydrates and glycoproteins. Anal. Chem. 77, 393-399. 

Ecker, R. C., de Martin, R., Steiner, G. E. and Schmid, J. A. (2004). Application of spectral imaging microscopy in cytomics and fluorescence resonance energy transfer (FRET) analysis. Cytometry A 59, 172-81. 

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