Protein Resilience and Fluorescent Protein Resistance to Photobleaching

Genetically Engineered Mesenchymal Stem Cells Stably Expressing Green Fluorescent Protein

Objective(s) Mesenchymal stem cells (MSCs) are nonhematopoietic stromal cells that are capable of differentiating into and contribute to the regeneration of mesenchymal tissues. Human mesenchymal stem cells (hMSCs) are ideal targets in cell transplantation and tissue engineering. Enhanced green fluorescent protein (EGFP) has been an important reporter gene for gene therapy. The aim of this stu...

Protonation, photobleaching, and photoactivation of yellow fluorescent protein (YFP 10C): a unifying mechanism.

Yellow fluorescent protein (YFP 10C) is widely used as a probe in biology, but its complex photochemistry gives rise to unusual behavior that requires fuller definition. Here we characterize the kinetics of protonation and reversible bleaching over time scales of picoseconds to hours. Stopped-flow and pressure-jump techniques showed that protonation of the fluorescent YFP(-) anion state is two-...

Green fluorescent protein chimeras to probe protein-protein interactions.

Reduced protein diffusion rate as revealed with fluorescence recovery after photobleaching of the green fluorescent protein in live cells

Fluorescence recovery after photobleaching measurements are performed to elucidate the mobility of the green fluorescent protein (GFP) in aqueous domains of Dictyostelium discoideum amoebae. A confocal detection scheme is adopted that allows one to record the recovery dynamics which originate exclusively from intracellular regions. In the confocal arrangement, favorable excitation intensity lev...

Reduced protein diffusion rate as revealed with fluorescence recovery after photobleaching of the green fluorescent protein in live cells

Fluorescence recovery after photobleaching measurements are performed to elucidate the mobility of the green fluorescent protein (GFP) in aqueous domains of Dictyostelium discoideum amoebae. A confocal detection scheme is adopted that allows one to record the recovery dynamics which originate exclusively from intracellular regions. In the confocal arrangement, favorable excitation intensity lev...