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The Journal of Cell Biology – November 2014

pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis
The Journal of Cell Biology – November 2014. doi:10.1083/jcb.201404107

Fluorescent proteins with pH-sensitive fluorescence are valuable tools for the imaging of exocytosis and endocytosis, fundamental processes in cells. The Aequorea green fluorescent protein (GFP) 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.

pHuji - Journal of Cell Biology

Yi Shen, Morgane Rosendale, Robert E Campbell & David Perrais

David Perrais, tel. 05 57 57 56 92