%0 Journal Article
%T Color transitions in coral's fluorescent proteins by site-directed mutagenesis
%A Nadya G Gurskaya
%A Alexander P Savitsky
%A Yurii G Yanushevich
%A Sergey A Lukyanov
%A Konstantin A Lukyanov
%J BMC Biochemistry
%D 2001
%I BioMed Central
%R 10.1186/1471-2091-2-6
%X Here we applied site-directed mutagenesis in order to investigate the structural background of color variety and possibility of shifting between different types of fluorescence. First, a blue-shifted mutant of cyan amFP486 was generated. Second, it was established that cyan and green emitters can be modified so as to produce an intermediate spectrum of fluorescence. Third, the relationship between green and yellow fluorescence was inspected on closely homologous green zFP506 and yellow zFP538 proteins. The following transitions of colors were performed: yellow to green; yellow to dual color (green and yellow); and green to yellow. Fourth, we generated a mutant of cyan emitter dsFP483 that demonstrated dual color (cyan and red) fluorescence.Several amino acid substitutions were found to strongly affect fluorescence maxima. Some positions primarily found by sequence comparison were proved to be crucial for fluorescence of particular color. These results are the first step towards predicting the color of natural GFP-like proteins corresponding to newly identified cDNAs from corals.Since the introduction of Green Fluorescent Protein (GFP) into biotechnology many attempts have been made to change its fluorescence color by means of mutagenesis. Finally, only two positions on protein sequence were demonstrated to considerably influence emission maximum [1,2,3]. First, a replacement of fluorophore's Tyr-66 with any aromatic residue (Trp, Phe, or His) results in a strong blue shift of emission up to ¦Ëmax = 442 nm [4, 5]. Second, a substitution S203Y,H leads to a red shift of fluorescence up to ¦Ëmax = 529 nm [6]. These blue and yellow mutants (called BFP and YFP respectively) proved to be extremely handy for multicolor labeling and FRET-based applications.Recently we have isolated several GFP-like proteins that determine fluorescent or non-fluorescent body color in corals Anthozoa [7,8,9]. Absorption-emission maxima of these proteins are distributed loosely along the waveleng
%U http://www.biomedcentral.com/1471-2091/2/6