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Cited by 0Receptor Mechanisms and SignalingCellular transport and secretionChemokine receptors and signaling

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A high-resolution analysis of arrestin2 interactions responsible for CCR5 endocytosis

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Petrovic, Ivana, Desai, Samit, Isaikina, Polina, Abiko, Layara Akemi, Spang, Anne, Grzesiek, Stephan·2026·eLife
DOI 10.7554/elife.106839.4

Preprint — published as 10.7554/elife.106839

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Abstract

Clathrin-mediated endocytosis (CME) is crucial for regulating G protein-coupled receptors (GPCRs) via phosphorylation-dependent arrestin interactions. Despite detailed structural knowledge on the arrestin interactions with phosphorylated tails of GPCRs, the interplay between receptor phosphorylation and arrestin coupling to the CME machinery is not well understood, in particular due to the weakness and dynamics of the individual molecular interactions. Here, we have characterized the interactions of arrestin2, which is activated by the phosphorylated C-terminus of the human chemokine receptor 5 (CCR5), with the main protein constituents of CME, namely clathrin and AP2, by solution NMR spectroscopy, biochemical, and cellular assays. The NMR analysis revealed that arrestin2 interacts weakly with clathrin through a single binding site, independent of arrestin2 activation. In contrast, the arrestin2-AP2 interaction is stronger, requires arrestin2 activation by the CCR5 phospho-tail, and depends quantitatively on its degree of phosphorylation. These in vitro results are corroborated by cellular assays, which show that the chemokine-induced formation of a long-lived CCR5-arrestin2 internalization complex depends strongly on the interaction of arrestin2 with AP2, but not with clathrin. Taken together, these findings provide quantitative, atom-scale insights on the first steps of CCR5 endocytosis.

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Metadata

Authors
Petrovic, I., Desai, S., Isaikina, P., Abiko, L. A., Spang, A., & Grzesiek, S.
Affiliations
  • University of Basel (CH)
  • Paul Scherrer Institute (CH)
Corresponding author
Grzesiek, Stephan
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2026-01-19
Volume
14
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.106839.4
OpenAlex
W4404087773
Cited by
0
References
3
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
Funders
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (31-201270, IZLIZ3-200298, 31-197779, 31-185127)
  • Fellowship for Excellence by the Biozentrum Basel International PhD Program
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Health Sciences › Medicine › Oncology › Chemokine receptors and signaling
Keywords
arrestin-2, CCR5, chemokine receptor 5, clathrin-mediated endocytosis, AP-2, receptor phosphorylation, G protein-coupled receptor internalization, solution NMR

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