Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning.
The cloning and expression of autofluorescent proteins in living matter, combined with modern imaging techniques, have thoroughly changed the world of bioscience. In particular, such proteins are widely used as genetically encoded labels to track the movement of proteins as reporters of cellular signals and to study protein-protein interactions by fluorescence resonance energy transfer (FRET). Their optical properties, however, are complex and it is important to understand these for the correct interpretation of imaging data and for the design of new fluorescent mutants. In this Minireview we start with a short survey of the field and then focus on the photo- and thermally induced dynamics of green and red fluorescent proteins. In particular, we show how fluorescence line narrowing and high-resolution spectral hole burning at low temperatures can be used to unravel the photophysics and photochemistry and shed light on the intricate electronic structure of these proteins.
Bonsma
S
S
Huygens and Gorlaeus Laboratories, Leiden University, P.O. Box 9504, 2300 RA Leiden, The Netherlands.
Purchase
R
R
Jezowski
S
S
Gallus
J
J
Könz
F
F
Völker
S
S
D001703:Biophysics / Q000379:methods*; D002627:Chemistry, Physical / Q000379:methods*; D004583:Electrons; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006358:Hot Temperature; D007834:Lasers; D008027:Light; D008164:Luminescent Proteins / Q000737:chemistry* / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D008968:Molecular Conformation; D009154:Mutation; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013053:Spectrophotometry / Q000379:methods*; D013696:Temperature; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C413663:fluorescent protein 583