O-Propargyl-puromycin, Acetate salt

Liu et al.[1] reported a non-radioactive alternative to analyze newly synthesized proteins in cell culture and whole organisms that is based on O-Propargyl-puromycin, an alkyne analog of puromycin. O-Propargyl-puromycin is cell-permeable and incorporates into the C-terminus of translating polypeptide chains thereby stopping translation. The resulting truncated C-terminal alkyne labeled proteins can subsequently be detected via Cu (I) -catalyzed click chemistry that offers the choice to introduce a Biotin group (via Azides of Biotin) for subsequent purification tasks or a fluorescent group (via Azides of fluorescent dyes) for subsequent microscopic imaging. In contrast to Azidohomoalanine (AHA) or Homopropargylgycine (HPG) based non-radioactive methionine analog-approaches, methionine free-medium is not required for O-Propargyl-purmoycin-based monitoring of nascent protein synthesis. Presolski et al.[4] and Hong et al.[5] provide a general protocol for Cu (I) -catalyzed click chemistry reactions that may be used as a starting point for the set up and optimization of individual assays.