Design of Photocaged Puromycin for Nascent Polypeptide Release and Spatiotemporal Monitoring of Translation |
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| Authors: | Florian Buhr Jörg Kohl‐Landgraf Susanne tom Dieck Cyril Hanus Deep Chatterjee Andreas Hegelein Prof Dr Erin M Schuman Prof Dr Josef Wachtveitl Prof Dr Harald Schwalbe |
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| Institution: | 1. Center for Biomolecular Magnetic Resonance, Institute of Organic Chemistry and Chemical Biology, Goethe‐Universit?t Frankfurt am Main, Max‐von‐Laue‐Stra?e 7, 60438 Frankfurt am Main (Germany);2. Institute of Physical and Theoretical Chemistry, Goethe‐Universit?t Frankfurt am Main, Max‐von‐Laue‐Stra?e 7, 60438 Frankfurt am Main (Germany);3. Department of Synaptic Plasticity, MPI for Brain Research, Max‐von‐Laue‐Stra?e 4, 60438 Frankfurt am Main (Germany) |
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| Abstract: | The antibiotic puromycin, which inhibits protein translation, is used in a broad range of biochemical applications. The synthesis, characterization, and biological applications of NVOC‐puromycin, a photocaged derivative that is activated by UV illumination, are presented. The caged compound had no effect either on prokaryotic or eukaryotic translation or on the viability of HEK 293 cells. Furthermore, no significant release of ribosome‐bound polypeptide chains was detected in vitro. Upon illumination, cytotoxic activity, in vitro translation inhibition, and polypeptide release triggered by the uncaging of NVOC‐puromycin were equivalent to those of the commercial compound. The quantum yield of photolysis was determined to be 1.1±0.2 % and the NVOC‐puromycin was applied to the detection of newly translated proteins with remarkable spatiotemporal resolution by using two‐photon laser excitation, puromycin immunohistochemistry, and imaging in rat hippocampal neurons. |
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| Keywords: | antibiotics inhibitors photoactivation protecting groups protein expression |
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