Anson, Francesca; Kanjilal, Pintu; Thayumanavan, S.; Hardy, Jeanne A. published the artcile< Tracking exogenous intracellular casp-3 using split GFP>, Reference of 2127-03-9, the main research area is green fluorescent protein exogenous intracellular caspase cytosol; apoptosis; caspase; caspase-3; intracellular protein delivery; nanogel; split GFP.
Cytosolic protein delivery promises diverse applications from therapeutics, to genetic modification and precision research tools. To achieve effective cellular and subcellular delivery, approaches that allow protein visualization and accurate localization with greater sensitivity are essential. Fluorescently tagging proteins allows detection, tracking and visualization in cellulo. However, undesired consequences from fluorophores or fluorescent protein tags, such as nonspecific interactions and high background or perturbation to native protein””s size and structure, are frequently observed, or more troublingly, overlooked. Distinguishing cytosolically released mols. from those that are endosomally entrapped upon cellular uptake is particularly challenging and is often complicated by the inherent pH-sensitive and hydrophobic properties of the fluorophore. Monitoring localization is more complex in delivery of proteins with inherent protein-modifying activities like proteases, transacetylases, kinases, etc. Proteases are among the toughest cargos due to their inherent propensity for self-proteolysis. To implement a reliable, but functionally silent, tagging technol. in a protease, we have developed a caspase-3 variant tagged with the 11th strand of GFP that retains both enzymic activity and structural characteristics of wild-type caspase-3. Only in the presence of cytosolic GFP strands 1-10 will the tagged caspase-3 generate fluorescence to signal a non-endosomal location. This methodol. facilitates easy screening of cytosolic vs. endosomally-entrapped proteins due to low probabilities for false pos. results, and further, allows tracking of the resultant cargo′s translocation. The development of this tagged casp-3 cytosolic reporter lays the foundation to probe caspase therapeutic properties, charge-property relationships governing successful escape, and the precise number of caspases required for apoptotic cell death.
Protein Science published new progress about Affinity tags Role: BSU (Biological Study, Unclassified), BIOL (Biological Study). 2127-03-9 belongs to class pyridine-derivatives, and the molecular formula is C10H8N2S2, Reference of 2127-03-9.