%0 Journal Article %A Mizukami, Shin %A Watanabe, Shuji %A Akimoto, Yuri %A Kikuchi, Kazuya %D 2012 %T No-Wash Protein Labeling with Designed Fluorogenic Probes and Application to Real-Time Pulse-Chase Analysis %U https://acs.figshare.com/articles/journal_contribution/No_Wash_Protein_Labeling_with_Designed_Fluorogenic_Probes_and_Application_to_Real_Time_Pulse_Chase_Analysis/2556244 %R 10.1021/ja208290f.s001 %2 https://acs.figshare.com/ndownloader/files/4199320 %K Fast quencher elimination %K fluorescence resonance energy transfer %K epidermal growth factor receptors %K cell surface %K Fluorogenic Probes %K intracellular proteins %K intracellular region %K probe %K fluorogenic %K FRET %X Small molecule labeling techniques for cellular proteins under physiological conditions are very promising for revealing new biological functions. We developed a no-wash fluorogenic labeling system by exploiting fluorescence resonance energy transfer (FRET)-based fluorescein-cephalosporin-azopyridinium probes and a mutant β-lactamase tag. Fast quencher elimination, hydrophilicity, and high resistance against autodegradation were achieved by rational refinement of the structure. By applying the probe to real-time pulse-chase analysis, the trafficking of epidermal growth factor receptors between cell surface and intracellular region was imaged. In addition, membrane-permeable derivatization of the probe enabled no-wash fluorogenic labeling of intracellular proteins. %I ACS Publications