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Genetically Encoded pH Sensor for Tracking Surface Proteins through Endocytosis

Authors

  • Anmol Grover,

    1. Department of Biological Sciences and Molecular Biosensor and Imaging Center, Carnegie Mellon University (USA)
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  • Dr. Brigitte F. Schmidt,

    1. Molecular Biosensor and Imaging Centre, Carnegie Mellon University (USA)
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  • Dr. Russell D. Salter,

    1. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, PA (USA)
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  • Dr. Simon C. Watkins,

    1. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, PA (USA)
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  • Dr. Alan S. Waggoner,

    1. Department of Biological Sciences and Molecular Biosensor and Imaging Center, Carnegie Mellon University (USA)
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  • Dr. Marcel P. Bruchez

    Corresponding author
    1. Department of Chemistry, Department of Biological Sciences and Molecular Biosensor and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213 (USA)
    • Department of Chemistry, Department of Biological Sciences and Molecular Biosensor and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213 (USA)
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  • This project was supported by grants from the National Center for Research Resources (5U54RR022241-08) and the National Institute of General Medical Sciences (8 U54 GM103529-08) from the National Institutes of Health. We would like to thank Jonathan W. Jarvik for the gift of stable ADRB2 cells, Yehuda Creeger for help with flow cytometry, and Haibing Teng for help with confocal microscopy. We would like to thank Lauren Ernst and Chris Szent-Gyorgyi for helpful discussions, and Kristen McConnell for assistance preparing the illustration. Microscopes used for the project were acquired under NIH grants 1S10RR024716 and 1S10RR026766.

Abstract

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Verkehrsüberwachung: Ein Tandemfarbstoff aus einem FRET-Akzeptor und einem fluorogenen Donor funktioniert als ratiometrischer pH-Indikator für Zelloberflächen, mit dem sich nach Internalisierung der Proteintransport während der Endozytose verfolgen lässt. Der Sensor wurde auf zweierlei Weise eingesetzt: zur Analyse der Agonist-abhängigen Internalisierung des β2-adrenergen Rezeptors (siehe Schema) und zur Untersuchung des direkten Antigentransfers von der Oberfläche zum Endosom zwischen dendritischen Zellen.

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