Imaging protein interactions with bioluminescence resonance energy transfer (BRET) in plant and mammalian cells and tissues Export

@article{citeulike:1367460, abstract = {Edited by J. Woodland Hastings, Harvard University, Cambridge, MA, and approved April 16, 2007 (received for review March 3, 2007)FRET is a well established method for cellular and subcellular imaging of protein interactions. However, FRET obligatorily necessitates fluorescence excitation with its concomitant problems of photobleaching, autofluorescence, phototoxicity, and undesirable stimulation of photobiological processes. A sister technique, bioluminescence resonance energy transfer (BRET), avoids these problems because it uses enzyme-catalyzed luminescence; however, BRET signals usually have been too dim to image effectively in the past. Using a new generation electron bombardment-charge-coupled device camera coupled to an image splitter, we demonstrate that BRET can be used to image protein interactions in plant and animal cells and in tissues; even subcellular imaging is possible. We have applied this technology to image two different protein interactions: (i) dimerization of the developmental regulator, COP1, in plant seedlings; and (ii) CCAAT/enhancer binding protein {alpha} (C/EBP{alpha}) in the mammalian nucleus. This advance heralds a host of applications for imaging without fluorescent excitation and its consequent limitations. 10.1073/pnas.0701987104}, author = {Xu, Xiaodong and Soutto, Mohammed and Xie, Qiguang and Servick, Stein and Subramanian, Chitra and von Arnim, Albrecht G. and Johnson, Carl H.}, citeulike-article-id = {1367460}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0701987104}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17551013}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17551013}, day = {5}, doi = {10.1073/pnas.0701987104}, journal = {PNAS}, keywords = {bret, interaction, protein}, month = {June}, pages = {0701987104+}, posted-at = {2007-06-06 08:48:18}, priority = {3}, title = {Imaging protein interactions with bioluminescence resonance energy transfer (BRET) in plant and mammalian cells and tissues}, url = {http://dx.doi.org/10.1073/pnas.0701987104}, year = {2007} }

TY - JOUR ID - citeulike:1367460 L3 - citeulike-article-id:1367460 N2 - Edited by J. Woodland Hastings, Harvard University, Cambridge, MA, and approved April 16, 2007 (received for review March 3, 2007)FRET is a well established method for cellular and subcellular imaging of protein interactions. However, FRET obligatorily necessitates fluorescence excitation with its concomitant problems of photobleaching, autofluorescence, phototoxicity, and undesirable stimulation of photobiological processes. A sister technique, bioluminescence resonance energy transfer (BRET), avoids these problems because it uses enzyme-catalyzed luminescence; however, BRET signals usually have been too dim to image effectively in the past. Using a new generation electron bombardment-charge-coupled device camera coupled to an image splitter, we demonstrate that BRET can be used to image protein interactions in plant and animal cells and in tissues; even subcellular imaging is possible. We have applied this technology to image two different protein interactions: (i) dimerization of the developmental regulator, COP1, in plant seedlings; and (ii) CCAAT/enhancer binding protein {alpha} (C/EBP{alpha}) in the mammalian nucleus. This advance heralds a host of applications for imaging without fluorescent excitation and its consequent limitations. 10.1073/pnas.0701987104 JF - PNAS TI - Imaging protein interactions with bioluminescence resonance energy transfer (BRET) in plant and mammalian cells and tissues SP - 0701987104 KW - bret KW - interaction KW - protein AU - Xu, Xiaodong AU - Soutto, Mohammed AU - Xie, Qiguang AU - Servick, Stein AU - Subramanian, Chitra AU - von Arnim, Albrecht AU - Johnson, Carl PY - 2007/06/05/ UR - http://dx.doi.org/10.1073/pnas.0701987104 ER -