期刊
CHEMBIOCHEM
卷 19, 期 23, 页码 2458-2464出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201800408
关键词
analytical chemistry; coumarin; light control; neuronal control; puromycin
资金
- DFG in graduate college CLIC
- state of Hessen
- DFG in the collaborative research center SFB902
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.
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