The use of FRET imaging microscopy to detect protein-protein interactions and protein conformational changes in vivo.
Intermolecular and intramolecular FRET between two spectrally overlapping green fluorescent protein variants fused to two different host proteins or at two different sites within the same protein offers a unique opportunity to monitor real-time protein-protein interactions or protein conformational changes. By using fluorescence digital imaging microscopy, one can visualize the location of green fluorescent proteins within a living cell and follow the time course of the changes in FRET corresponding to cellular events at a millisecond time resolution. The observation of such dynamic molecular events in vivo provides vital insight into the action of biological molecules.
Truong
K
K
Division of Molecular and Structural Biology, Ontario Cancer Institute, Toronto, Canada.
Ikura
M
M
D020013:Calcium Signaling; D010450:Endopeptidases / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D046911:Macromolecular Substances; D008856:Microscopy, Fluorescence / Q000379:methods*; D010766:Phosphorylation; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
D008164:Luminescent Proteins; D046911:Macromolecular Substances; D011506:Proteins; D049452:Green Fluorescent Proteins; D010450:Endopeptidases