pHuji,a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis |
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| Authors: | Yi Shen Morgane Rosendale Robert E Campbell David Perrais |
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| Institution: | 1.Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada;2.University of Bordeaux, and 3.Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux, France |
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| 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. |
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