In vivo detection of protein-protein interaction in plant cells using BRET.
The emerging technique of bioluminescence resonance energy transfer (BRET) allows us to detect protein interactions in live cells and in real time, thus providing a new window into cellular signal transduction processes. We present experimental protocols for expressing fusion proteins between luciferase and fluorescent proteins that are the basis for BRET measurement, as well as for measuring and imaging BRET in a variety of cell types. Despite our focus on plant cells, the techniques described here are easily adaptable to other cell systems that have yet to benefit from the BRET technique.
Subramanian
Chitra
C
Department of Botany, The University of Tennessee, Knoxville, USA.
Xu
Yao
Y
Johnson
Carl Hirschie
CH
von Arnim
Albrecht G
AG
D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D004735:Energy Transfer; D008156:Luciferases / Q000737:chemistry; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D019697:Onions / Q000737:chemistry / Q000235:genetics; D010940:Plant Proteins / Q000032:analysis* / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015398:Signal Transduction; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D008156:Luciferases