Monitoring newly synthesized proteins is becoming increasingly important to characterize proteome composition in regulatory networks. Puromycin is a peptidyl transfer inhibitor, widely used in cell biology for tagging newly synthesized proteins. Here, we report synthesis and application of an optimized puromycin carrying a photolabile protecting group as a powerful tool for tagging nascent proteins with high spatiotemporal resolution. The photocaged 7‐N,N‐(diethylaminocumarin‐4‐yl)‐methoxycarbonyl‐puromycin (DEACM‐puromycin) was synthesized and compared with the previously developed 6‐nitroveratryloxycarbonyl puromycin (NVOC‐puromycin). The photochemical behavior as well as the effectiveness in controlling puromycylation in living hippocampal neurons using two‐photon excitation is superior to the previously used NVOCpuromycin. We further report on the application of light‐controlled puromycylation to visualize new translated proteins in neurons.
The cover feature picture shows a novel photocaged antibiotic puromycin for the spatiotemporal control and monitoring of nascent polypeptide chain translation. Following two‐photon uncaging, the photoreleased puromycin is attached to the nascent polypeptide chain and can be detected via antibody as a puromycylation signal in the small region of the cell body of an illuminated hippocampal neuron. Light‐controlled tagging of nascent proteins with such high incorporation efficiency and spatial resolution will expand the chemical biology tools for live quantification of protein translation. More information can be found in the full paper by H. Schwalbe et al. on page 2458 in Issue 23, 2018 (DOI: 10.1002/cbic.201800408).
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