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Mutants of Discosoma red fluorescent protein with a GFP‐like chromophore
Author(s) -
Wiehler Jens,
von Hummel Julia,
Steipe Boris
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)02365-6
Subject(s) - green fluorescent protein , chromophore , fluorescence , fluorophore , chemistry , mutant , biophysics , fluorescent protein , photochemistry , biochemistry , biology , gene , optics , physics
The green fluorescent protein (GFP)‐homologous red fluorescent protein (RFP) from Discosoma (drFP583) which emits bright red fluorescence peaking at 583 nm is an interesting novel genetic marker. We show here that RFP maturation involves a GFP‐like fluorophore which can be stabilized by point mutations selected from a randomly mutated expression library. By homology modeling, these point mutations cluster near the imidazolidinone ring of the chromophore. Exciting the GFP‐like absorption band in the mutant proteins produces both green and red fluorescence. Upon unfolding and heating, the absorption spectrum of the RFP chromophore slowly becomes similar to that of the GFP chromophore. This can be interpreted as a covalent modification of the GFP chromophore in RFP that appears to occur in the final maturation step.