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Perkins, L.A.* ; Yan, Q.* ; Schmidt, B.F.* ; Kolodieznyi, D.* ; Saurabh, S.* ; Larsen, M.B.* ; Watkins, S.C.* ; Kremer, L.S. ; Bruchez, M.P.*

Genetically targeted ratiometric and activated pH indicator complexes (TRApHIC) for receptor trafficking.

Biochemistry 57, 861-871 (2018)
Postprint DOI PMC
Open Access Green
Fluorescent protein-based pH sensors are useful tools for measuring protein trafficking through pH changes associated with endo- and exocytosis. However, commonly used pH-sensing probes are ubiquitously expressed with their protein of interest throughout the cell, hindering our ability to focus on specific trafficking pools of proteins. We developed a family of excitation ratiometric, activatable pH responsive tandem dyes, consisting of a pH sensitive Cy3 donor linked to a fluorogenic malachite green acceptor. These cell-excluded dyes are targeted and activated upon binding to a genetically expressed fluorogen-activating protein and are suitable for selective labeling of surface proteins for analysis of endocytosis and recycling in live cells using both confocal and superresolution microscopy. Quantitative profiling of the endocytosis and recycling of tagged beta 2-adrenergic receptor (B2AR) at a single-vesicle level revealed differences among B2AR agonists, consistent with more detailed pharmacological profiling.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Red Fluorescent Protein; Beta(2)-adrenergic Receptor; Surface-proteins; Intracellular Ph; Cell; Probes; Internalization; Endocytosis; Microscopy; Pathways
Sprache englisch
Veröffentlichungsjahr 2018
HGF-Berichtsjahr 2018
ISSN (print) / ISBN 0006-2960
e-ISSN 1520-4995
Zeitschrift Biochemistry
Quellenangaben Band: 57, Heft: 5, Seiten: 861-871 Artikelnummer: , Supplement: ,
Verlag American Chemical Society (ACS)
Verlagsort Washington
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Human Genetics (IHG)
POF Topic(s) 30501 - Systemic Analysis of Genetic and Environmental Factors that Impact Health
Forschungsfeld(er) Genetics and Epidemiology
PSP-Element(e) G-500700-001
DOI 10.1021/acs.biochem.7b01135
WOS ID WOS:000424723300049
Scopus ID 85041456948
PubMed ID 29283245
Erfassungsdatum 2018-03-12
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