Arnaud Gautier has just published how The Twinkle Factory may help to differentially label cell-surface and intracellular membrane protein based on the chemical-genetic fluorescent marker FAST. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, including flow cytometry, with no imaging!

Check Bioconjugate Chem.: Fluorogenic Probing of Membrane Protein Trafficking. Bioconjug. Chem. (2018) 29(6), 1823-1828

Abstract

Methods to differentially label cell-surface and intracellular membrane proteins are indispensable for understanding their function and the regulation of their trafficking. We present an efficient strategy for the rapid and selective fluorescent labeling of membrane proteins based on the chemical-genetic fluorescent marker FAST (fluorescence-activating and absorption-shifting tag). Cell-surface FAST-tagged proteins could be selectively and rapidly labeled using fluorogenic membrane-impermeant 4-hydroxybenzylidene rhodanine (HBR) analogs. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, and opens exciting prospects for the high-throughput screening of small molecules able to restore disease-related trafficking defects.

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