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J Cell Biol. 2014 Nov 10;207(3):419-32. doi: 10.1083/jcb.201404107.

pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis.

Author information

1
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
2
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.
3
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada robert.e.campbell@ualberta.ca david.perrais@u-bordeaux.fr.
4
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.

Abstract

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.

PMID:
25385186
PMCID:
PMC4226733
DOI:
10.1083/jcb.201404107
[Indexed for MEDLINE]
Free PMC Article

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