Journal
JOURNAL OF CELL BIOLOGY
Volume 207, Issue 3, Pages 419-432Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.201404107
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Funding
- Natural Sciences and Engineering Research Council of Canada
- Fondation Recherche Medicale
- Centre National de la Recherche Scientifique (Interface program)
- Agence Nationale pour la Recherche [CaPeBlE ANR-12-BSV5-005]
- Alberta Ingenuity Scholarship
- University of Bordeaux
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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. 2 adrenergic receptor. These experiments reveal that the two receptors are differentially sorted at the time of endocytic vesicle formation.
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