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...
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| Veröffentlicht in: | The Journal of cell biology 2014-11, Vol.207 (3), p.419-432 |
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| creator | Shen, Yi Rosendale, Morgane Campbell, Robert E Perrais, David |
| description | 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. |
| doi_str_mv | 10.1083/jcb.201404107 |
| format | Article |
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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. 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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. 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| subjects | Animals Cellular Biology Clathrin-Coated Vesicles - metabolism Directed Molecular Evolution Endocytosis Exocytosis Fluorescence Hydrogen-Ion Concentration Life Sciences Luminescent Proteins - chemistry Luminescent Proteins - genetics Luminescent Proteins - metabolism Mice Microscopy, Fluorescence Mutation NIH 3T3 Cells Protein Transport Proteins Red Fluorescent Protein |
| title | pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis |
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