1. Fluorogenic Probing of Membrane Protein Trafficking.
- Author
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Li C, Mourton A, Plamont MA, Rodrigues V, Aujard I, Volovitch M, Le Saux T, Perez F, Vriz S, Jullien L, Joliot A, and Gautier A
- Subjects
- Benzylidene Compounds analysis, Cell Membrane chemistry, Fluorescent Dyes analysis, HEK293 Cells, Humans, Luminescent Proteins analysis, Luminescent Proteins metabolism, Membrane Proteins analysis, Microscopy, Fluorescence methods, Protein Transport, Rhodanine analysis, Rhodanine metabolism, Red Fluorescent Protein, Benzylidene Compounds metabolism, Cell Membrane metabolism, Fluorescent Dyes metabolism, Membrane Proteins metabolism, Rhodanine analogs & derivatives
- 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.
- Published
- 2018
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