pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis.
Fluorescent proteins with pH-sensitive fluorescence are valuable tools for the imaging of exocytosis and endocytosis. The Aequorea green fluorescent protein mutant superecliptic pHluorin (SEP) is particularly well suited to these applications. Here we describe pHuji, a red fluorescent protein with a pH sensitivity that approaches that of SEP, making it amenable for detection of single exocytosis and endocytosis events. To demonstrate the utility of the pHuji plus SEP pair, we perform simultaneous two-color imaging of clathrin-mediated internalization of both the transferrin receptor and the β2 adrenergic receptor. These experiments reveal that the two receptors are differentially sorted at the time of endocytic vesicle formation.
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
Yi
Y
Shen
University of Bordeaux and Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France University of Bordeaux and Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France.
Morgane
M
Rosendale
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada robert.e.campbell@ualberta.ca david.perrais@u-bordeaux.fr.
Robert E
RE
Campbell
University of Bordeaux and Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France University of Bordeaux and Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France robert.e.campbell@ualberta.ca david.perrais@u-bordeaux.fr.
David
D
Perrais
D000818:Animals; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D019020:Directed Molecular Evolution; D004705:Endocytosis; D005089:Exocytosis; D006863:Hydrogen-Ion Concentration; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D021381:Protein Transport; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein