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Abstract
The GFP-based superecliptic pHluorin (SEP) enables detection of exocytosis and endocytosis, but its performance has not been duplicated in red fluorescent protein scaffolds. Here we describe “semisynthetic” pH-sensitive protein conjugates with organic fluorophores, carbofluorescein, and Virginia Orange that match the properties of SEP. Conjugation to genetically encoded self-labeling tags or antibodies allows visualization of both exocytosis and endocytosis, constituting new bright sensors for these key steps of synaptic transmission.
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1 University of Bordeaux, Bordeaux, France; Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, Bordeaux, France
2 National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, MD, USA
3 Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA
4 University of Bordeaux, Bordeaux, France; Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, Bordeaux, France; Bordeaux Imaging Center, UMS 3420 CNRS, Université de Bordeaux, US 4 INSERM, Bordeaux, France