Coordinate-based co-localization-mediated analysis of arrestin clustering upon stimulation of the C–C chemokine receptor 5 with RANTES/CCL5 analogues

G protein-coupled receptor activation and desensitization leads to recruitment of arrestin proteins from cytosolic pools to the cell membrane where they form clusters difficult to characterize due to their small size and further mediate receptor internalization. We quantitatively investigated cluste...

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Bibliographic Details
Published in:Histochemistry and cell biology 2014-07, Vol.142 (1), p.69-77
Main Authors: Tarancón Díez, Laura, Bönsch, Claudia, Malkusch, Sebastian, Truan, Zinnia, Munteanu, Mihaela, Heilemann, Mike, Hartley, Oliver, Endesfelder, Ulrike, Fürstenberg, Alexandre
Format: Article
Language:English
Subjects:
Animals
Arrestins - analysis
Arrestins - metabolism
Biochemistry
Biomedical and Life Sciences
Biomedicine
Cattle
Cell Biology
Cells
Cells, Cultured
Chemokine CCL5 - chemistry
Chemokine CCL5 - pharmacology
Chemokines
CHO Cells
Cricetulus
Developmental Biology
Membranes
Microscopy
Microscopy, Confocal
Microscopy, Fluorescence
Original Paper
Protein Transport - drug effects
Proteins
Receptors, CCR5 - agonists
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Summary:G protein-coupled receptor activation and desensitization leads to recruitment of arrestin proteins from cytosolic pools to the cell membrane where they form clusters difficult to characterize due to their small size and further mediate receptor internalization. We quantitatively investigated clustering of arrestin 3 induced by potent anti-HIV analogues of the chemokine RANTES after stimulation of the C–C chemokine receptor 5 using single-molecule localization-based super-resolution microscopy. We determined arrestin 3 cluster sizes and relative fractions of arrestin 3 molecules in each cluster through image-based analysis of the localization data by adapting a method originally developed for co-localization analysis from molecular coordinates. We found that only classical agonists in the set of tested ligands were able to efficiently recruit arrestin 3 to clusters mostly larger than 150 nm in size and compare our results with existing data on arrestin 2 clustering induced by the same chemokine analogues.
ISSN:0948-6143
1432-119X
DOI:10.1007/s00418-014-1206-1