Group of metabolic bases of pathology
The group was established in 2018.
The team of the group is engaged in the development of genetically encoded biosensors for imaging the dynamics of various biological processes in living systems in real time. Tools of this kind have gained wide popularity for solving various problems in biology and medicine. Such a biosensor is a protein molecule, which, as a rule, consists of two functional parts: sensory and fluorescent. Fluorescent proteins are used as a fluorescent domain. As the sensory part of the biosensor, natural domains of proteins that perform various functions in cells are most often used. The principle of operation of such a biosensor is based on the fact that under the influence of a specific stimulus, for example, with specific binding of a ligand, the sensory domain undergoes conformational changes that are transmitted to the fluorescent protein as part of the construct, which causes changes in spectral characteristics. Thus, the dynamics of changes in the fluorescent signal in a system reflects the dynamics of changes in the studied biological parameter.
This approach to the visualization of biological processes is becoming an integral part of modern research. Such methods are universal and applicable for use in a wide variety of biological objects. Using fluorescent biosensors, the scientific group is engaged in the in vivo study of physiological and pathological processes.
Currently, the group is working on in vivo studies of the dynamics of intracellular processes in brain tissue during the development of ischemic stroke in rodents. Another model object of the group is zebrafish D. rerio. Using this object, the group is engaged in the study of inflammatory processes caused by various factors.
- development of biosensors for registration of hypohalous acids and their derivatives;
- development of biosensors for studying the metabolism of fatty acids;
- development of biosensors for registration of hydrogen sulfide H2S;
- study of the key parameters of brain cells in the early stages of the development of ischemic stroke;
- study of redox processes in tissues in inflammatory reactions.
The ratio of oxidized and reduced forms of glutathione (GSSG/2GSH) is one of the most important cellular parameter. This parameter plays a paramount role in the reactions of the thiol-disulfide metabolism of the cell; many pathological processes are associated with impaired regulation of the GSSG/2GSH balance. Our team developed the Grx1-roCherry biosensor based on the red fluorescent protein. Depending on the redox state of the glutathione pool, Grx1-roCherry biosensor changes the fluorescent signal. This biosensor has been successfully tested using multiparameter microscopy in combination with various spectrally different biosensors in modeling different pathological conditions of cells.
Shokhina AG, Kostyuk AI, Ermakova YG, Panova AS, Staroverov DB, Egorov ES, Baranov MS, van Belle GJ, Katschinski DM, Belousov VV, Bilan DS. Red fluorescent redox-sensitive biosensor Grx1-roCherry. Redox Biology. 2019; 21: 101071.
|Dmitry Bilan, Ph.D.||firstname.lastname@example.org, |
|Alexandra Kokova (Demidovich)||j. r. email@example.com|
|Dar'ya Kotova||j. r. firstname.lastname@example.org, |
|Arina Shokhina||j. r. email@example.com, |
|Alexander Kostyuk||PhD firstname.lastname@example.org, |
|Anastasiya Panova||PhD email@example.com, |
Genetically encoded indicator Grx1-roCherry based on red fluorescent protein for detection of glutathione redox status
In collaboration with Laboratory of molecular technologies
We developed a genetically encoded biosensor for registration the redox state of the glutathione pool (2GSH/GSSG ratio) based on the red fluorescent protein mCherry. The structure of the fluorescent protein contains a pair of redox-active cysteines that are involved in the thiol-disulfide exchange reactions of intracellular systems. The fluorescent signal of the biosensor reflects the redox state of glutathione in the studied system due to the spectral difference between the oxidized and reduced forms of the protein. Human glutaredoxin-1 was added to the structure to improve the kinetic properties of the biosensor. Grx1-roCherry biosensor is a reliable tool for monitoring changes in the 2GSH/GSSG ratio in real time in various biological systems, including a combination with spectrally different versions in a multiparameter imaging mode.
- (2018). Red fluorescent redox-sensitive biosensor Grx1-roCherry. Redox Biol 21, 101071