Yellow fluorescent protein ( Eng. Yellow Fluorescent Protein ) - a genetic mutant form of green fluorescent protein ( GFP ), isolated from equus jellyfish Aequorea victoria . It is characterized by a maximum absorption at 514 nm and a maximum fluorescence at 527 nm. Widely used as a fluorescent label in cell and molecular biology to study the expression of cellular proteins. In addition to it, a series of other mutant forms of GFP was developed, such as blue, cyanic, etc. [1] .
Application for FRET
The absorption spectrum of YFP quite strongly overlaps with the fluorescence spectrum of CFP ( English Cyan Fluorescent Protein ), so these two fluorophores are used to create biosensors based on the FΓΆrster energy transfer (FRET) phenomenon. Such sensors are used to detect certain events occurring in living cells. In particular, enzyme activity can be determined in this way.
As a rule, the molecule of such a sensor includes 4 domains [2] :
- a protein that is a sensor carrier (it can be almost any protein);
- fluorophore CFP ;
- a domain that changes its structure in response to a specific effect (for example, phosphorylation );
- fluorophore YFP .
When such a sensor is irradiated with a laser with a wavelength that excites only CFP (for example, 440 nm β almost does not excite YFP , but strongly excites CFP ), one can observe the fluorescence of both fluorophores. After the corresponding effect on domain 3, the sensor structure changes, as a result of which the YFP fluorophore moves away from the CFP and the FΓΆrster transfer efficiency decreases (depending on how far the fluorophores were distant from each other). As a result, the fluorescence intensity of CFP increases, while YFP decreases. Thus, the ratio of YFP fluorescence to CFP fluorescence can be used to quantify the change in biosensor conformation .
Notes
- β Uemura, T .; Kim, H .; Saito, C .; Ebine, K .; Ueda, T .; Schulze-Lefert, P .; Nakano, A. Qa-SNAREs localized to the trans-Golgi network regulate multiple transport pathways and extracellular disease resistance in plants // Proceedings of the National Academy of Sciences of the United States of America : journal. - 2012. - Vol. 109 , no. 5 . - P. 1784-1789 . - DOI : 10.1073 / pnas . 1115146109 . - PMID 22307646 .
- β Access: Midzone activation of aurora B in anaphase produces an intracellular phosphorylation gradient: Nature
See also
- Green fluorescent protein
- Roger Tsien
- Fluorescence in biological research
Bibliography
- Miyawaki A., Llopis J., Heim R., et al . Fluorescent indicators for Ca2 + based on green fluorescent proteins and calmodulin (English) // Nature: journal. - 1997 .-- August ( vol. 388 , no. 6645 ). - P. 882β887 . - DOI : 10.1038 / 42264 . - PMID 9278050 .