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Spatiotemporal control of puromycin application with UV illumination

February 2015. Local synthesis of proteins near neuronal synapses is thought to be one of the key mechanisms involved in the cellular expression of learning and memory. The spatiotemporal monitoring of protein synthesis and the identification of these newly synthesized proteins is therefore a task of major importance. 

The antibiotic puromycin, an inhibitor of protein translation, is used in a broad range of biochemical applications. A team of Frankfurt scientists recently synthesized a photocaged derivative of puromycin that can be activated by UV illumination. This photocaged NVOC-puromycin will be of great use for a wide range of biological applications as it allows precise control of the location and timing of puromycin application. The scientists describe the synthesis, characterization and biological applications of NVOC-puromycin in the latest issue of the journal Angewandte Chemie International Edition. The team demonstrated photoinducible toxicity to eukaryotic cells, as well as the inhibition of protein translation in vitro in prokaryotic and eukaryotic cell-free expression systems. The scientists further showed that triggered polypeptide chain release can be achieved in vitro and also demonstrated the use of NVOC-puromycin to spatially control the release and incorporation of puromycin into living cells.

Puromycylation is a powerful mechanism for tagging nascent proteins. The demonstrated illumination-dependent spatiotemporal control of puromycin application in combination with living neurons promises an exciting approach for the study of localized protein synthesis near neuronal synapses and other systems. More ...


Contact:
Prof. Dr. Harald Schwalbe
Institute of Organic Chemistry and Chemical Biology
Center for Biomolecular Magnetic Resonance (BMRZ)
Goethe University Frankfurt
schwalbe@nmr.uni-frankfurt.de

 

Full publication:
Florian Buhr, Jçrg Kohl-Landgraf, Susanne tom Dieck, Cyril Hanus, Deep Chatterjee, Andreas Hegelein, Erin M. Schuman, Josef Wachtveitl, and Harald Schwalbe (2015) Design of photocaged puromycin for nascent polypeptide release and spatiotemporal monitoring of translation. Angew. Chem. Int. Ed. 54:Published online: 4 Feb 2015. More ...