C-Terminal Labeling of Proteins Using Fluorescently Conjugated Puromycin Derivatives
A novel C-terminal labeling technology has been developed using fluorescently conjugated puromycin derivatives in cell free extracts. It is an easy to handle, rapid and low cost method for protein research. The labeling process requires only the addition of a labeling reagent to the translation mixture. Puromycin is conjugated with a fluorophore that is incorporated into a protein at the C-terminus, thus linking the fluorophore to the protein. Recent advances in cell-free protein expression combined with the puromycin analogue C-terminal labeling technology resulted in novel applications, such as single molecule imaging, Kd determination by fluorescence cross-correlation spectroscopy (FCCS) among others. C-terminal labeling can be used for the introduction of a fluorophore and many other types of molecules such as affinity tags. This technology can be used for library screening, proteomics analysis and biosensors.