Abstract
Characterization of the chemical environment, movement, trafficking and interactions of proteins in live cells is essential to understanding their functions. Labeling protein with functional molecules is a widely used approach in protein research to elucidate the protein location and functions both in vitro and in live cells or in vivo. A peptide or a protein tag fused to the protein of interest and provides the opportunities for an attachment of small molecule probes or other fluorophore to image the dynamics of protein localization. Here we reviewed the recent development of no-wash small molecular probes for photoactive yellow protein (PYP-tag), by the means of utilizing a quenching mechanism based on the intramolecular interactions, or an environmental-sensitive fluorophore. Several fluorogenic probes have been developed, with fast labeling kinetics and cell permeability. This technology allows quick live-cell imaging of cell-surface and intracellular proteins without a wash-out procedure.
Highlights
Proteins account for some of the largest and most important molecules in living organism, and function through interaction with various biomolecules
fluorescent protein (FP) has been extensively used in protein labeling, because of its one domain structure which makes it easy and feasible to fuse into the target protein with no interference and dramatic change to the native protein functionalities, and owing to its compact structure [4,5,6]
The protein labeling system havebut no their fluorescence because of the intramolecular association association the fluorophores, fluorescence will upon be restored upon the between thebetween fluorophores, but their fluorescence intensity willintensity be restored the dissociation between thebetween fluorophores, but derivative their fluorescence intensity will be restored upon the dissociation dissociation coumarin its linked fluorescein
Summary
Proteins account for some of the largest and most important molecules in living organism, and function through interaction with various biomolecules. [1,7] Another useful protein labeling methodology is to label fusion protein with small molecule probes. Added to a culture of growing cells, and the probe can go across the cell membrane and bind the fusion peptide or protein to finish protein labeling(Figure (Figure1)1)[9]. The small molecule should be cell and and non-toxic to the the receptor protein. BLDisadvantage of this method is the free small molecule probemolecule needs to be removed the cells tags [20]. There is great to develop demanding to develop probesfor with requirement for any in live protein labeling probes protein with nolabeling requirement anyno washing procedure inwashing live cell procedure imaging. The fluorescent probethe canbackground largely minimize the the background and protein tag.
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