Probing protein-protein Interactions with FRET-FLIM.
The quantification of molecular interactions or conformational changes can conveniently be studied by using Förster Resonance Energy Transfer (FRET) as a spectroscopic ruler. The FRET phenomenon describes the transfer of energy from a donor to an acceptor molecule, if they are in close proximity (<10 nm). The most straightforward method to measure FRET is Fluorescence Lifetime Imaging Microscopy (FLIM). In this chapter, we will describe an application of FRET using FLIM to monitor the hexamer formation of CrFP/eYFP-labeled Arabidopsis thaliana cell division cycle protein (AtCDC48) expressed in plant protoplasts.
Bücherl
Christoph
C
Laboratory of Biochemistry, Wageningen University, Wageningen, Netherlands.
Aker
José
J
de Vries
Sacco
S
Borst
Jan Willem
JW
D000074183:ATPases Associated with Diverse Cellular Activities; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D015870:Gene Expression; D008856:Microscopy, Fluorescence / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods*; D011523:Protoplasts / Q000378:metabolism* / Q000648:ultrastructure; D014162:Transfection
D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D000074183:ATPases Associated with Diverse Cellular Activities; C525275:CDC48A protein, Arabidopsis