UNIGE document Scientific Article
previous document  unige:43123  next document
add to browser collection
Title

Quantitative morphological analysis of arrestin2 clustering upon G protein-coupled receptor stimulation by super-resolution microscopy

Authors
Malkusch, Sebastian
Endesfelder, Ulrike
Heilemann, Mike
show hidden authors show all authors [1 - 9]
Published in Journal of Structural Biology. 2013, vol. 184, no. 2, p. 329-34
Abstract Clustering of arrestins upon G protein-coupled receptor stimulation is a phenomenon that is well-known but difficult to describe quantitatively due to the size of the clusters close to the diffraction limit of visible light. We introduce a general method to quantitatively investigate the clustering of arrestin following stimulation of the C-C chemokine receptor 5 (CCR5) using single-molecule super-resolution imaging and coordinate and image-based cluster analysis. We investigated the effect of potent anti-HIV ligands of CCR5 with different pharmacological profiles on arrestin2 cluster formation and found that only the ligands capable of inducing CCR5 internalization induced arrestin2 recruitment and clustering. We further demonstrate that the fraction of arrestin2 molecules found in clusters larger than 100nm correlates with the magnitude of ligand-induced CCR5 internalization, but not with G protein activation, indicating that recruitment of arrestin2 to CCR5 is independent of G protein activation. Pre-treatment of the cells with the drug cytochalasin D, which blocks actin polymerization, led to the formation of larger clusters, whereas the inhibitor of microtubule polymerization nocodazole had little effect on arrestin2 recruitment, suggesting an active role of actin in the organization and dynamics of these aggregates.
Keywords Actin Cytoskeleton/drug effects/metabolismAnimalsArrestins/metabolismCHO CellsCattleChemokine CCL5/pharmacology/physiologyChemokines, CC/pharmacologyCricetinaeCricetulusCytochalasin D/pharmacologyGreen Fluorescent Proteins/metabolismMicroscopy, FluorescenceNocodazole/pharmacologyProtein TransportReceptors, CCR5/metabolismRecombinant Fusion Proteins/metabolismSingle-Domain Antibodies/chemistryTubulin Modulators/pharmacology
Identifiers
PMID: 24091038
Full text
Article (Published version) (1.6 MB) - document accessible for UNIGE members only Limited access to UNIGE
Structures
Research group Hiv (835)
Citation
(ISO format)
TRUAN, Zinnia et al. Quantitative morphological analysis of arrestin2 clustering upon G protein-coupled receptor stimulation by super-resolution microscopy. In: Journal of Structural Biology, 2013, vol. 184, n° 2, p. 329-34. https://archive-ouverte.unige.ch/unige:43123

171 hits

0 download

Update

Deposited on : 2014-12-11

Export document
Format :
Citation style :