![]() Being protein molecules, they are non-invasive and can be targeted to different cell types or even subcellular compartments, and, unlike chemical dyes, they are not prone to leaking from the cells during imaging. Unlike traditional analytical approaches, GEFIs provide the following advantages. Therefore, GEFIs convert biochemical events into macroscopic visible signals that can be detected using standard optical equipment. In general, GEFIs can be described as chimeric constructions based on at least one FP, the optical properties of which depend on a cellular parameter of interest. Īn extremely promising direction in modern research is the development of genetically encoded fluorescent indicators (GEFIs) based on aFPs that can be used to visualize and quantify enzymatic activity and conformational state of proteins as well as changes in the concentrations of particular molecules or biophysical parameters in vivo, including living cells, tissues, or whole organisms. Owing to their increasing scientific demand, different types of aFPs with optimized parameters-such as fluorescence intensity, maturation rate, phototoxicity, and oligomeric state-have been engineered using molecular biology methods. Many modern analytical techniques exploit the unique nature of these macromolecules in order to directly visualize structures and processes in living cells and organisms. Therefore, they can be expressed in different cellular systems maintaining their optical properties. ![]() The main reason for the wide applicability of aFPs is their ability to auto-catalytically form chromophores without requiring any additional factors. To date, a wide color variety of fluorescent proteins has been developed from different organisms, including representatives of other species such as Anthozoa, copepods, and even chordates. The first protein from this group called green fluorescent protein (GFP) was isolated from the jellyfish Aequorea victoria in 1962 and subsequently characterized. In this review, we highlight the basic principles of such sensors, the history of their creation, and a complete classification of the available biosensors.Īt present, autofluorescent proteins (aFPs) have become indispensable tools in many biological and medical studies. Conformational rearrangements of the sensory domain associated with ligand interaction or changes in the cellular parameter are transferred to the cpFP, changing the chromophore environment. One of the common principles of creating genetically encoded biosensors is based on the integration of a cpFP into a flexible region of a sensory domain or between two interacting domains, which are selected according to certain characteristics. Such a structure imparts greater mobility to the FP than that of the native variant, allowing greater lability of the spectral characteristics. In circularly permuted FPs (cpFPs), the original N- and C-termini are fused using a peptide linker, while new termini are formed near the chromophore. ![]() The circular permutation of single FPs led to the development of an extensive class of biosensors that allow the monitoring of many intracellular events. Genetically encoded biosensors based on fluorescent proteins (FPs) are a reliable tool for studying the various biological processes in living systems.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |