- Journal of Siberian Federal University. Biology: 2010, Vol. 3, Issue 4
- Eremeeva, Elena V.; Frank, Ludmila A.; Markova, Svetlana V.; Vysotski, Eugene
- Contact information
- Eremeeva, Elena V. : Institute of Biophysics SB RAS , Russia, 660036, Krasnoyarsk, Akademgorodok; Frank, Ludmila A. : Institute of Biophysics SB RAS Siberian Federal University , Russia, 660036, Krasnoyarsk, Akademgorodok Russia, 660041, Krasnoyarsk, Svobodny av., 79 , e-mail: ; Markova, Svetlana V. : Institute of Biophysics SB RAS Siberian Federal University , Russia, 660036, Krasnoyarsk, Akademgorodok Russia, 660041, Krasnoyarsk, Svobodny av., 79; Vysotski, Eugene : Institute of Biophysics SB RAS , Russia, 660036, Krasnoyarsk, Akademgorodok
- cgreGFP; obelin; C-end fusing; cgreGFP
Ca2+-regulated photoproteins genetically fused with biospecific polipeptides have been extensively used in intracellular Ca2+ measurements and in the development of binding assays. Gene fusions to aequorin have been limited to its N-terminus, since as previous studies indicated the protein loses bioluminescent activity upon modification of its C-terminus. To investigate this in regard to another photoprotein - obelin (OL) the one was elongated at its C-terminus with tyrosine (Y) and then fused with green fluorescent protein Clytia gregaria (cgreGFP) through the flexible 31 aa linker. Both proteins (OL-Y and OL-cgreGFP) were isolated and investigated. The OL-Y was found to form a stable photoprotein comlex, possessing 75 % of WT-OL bioluminescent activity. OL-cgreGFP activity preserves 46 % of WT-OL activity and demonstrates an effective resonance energy transfer, where OL-partner and cgreGFP-partner are energy donor and acceptor respectively. Thus, it was shown that the labels on the base of Ca2+-regulated photoprotein obelin may be obtained by fusing with biospecific polypeptides regardless its termini.
- Paper at repository of SibFU
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