PT  - JOURNAL ARTICLE
AU  - M. Florencia Sánchez
AU  - Marina S. Dietz
AU  - Ulrike Müller
AU  - Julian Weghuber
AU  - Karl Gatterdam
AU  - Ralph Wieneke
AU  - Mike Heilemann
AU  - Peter Lanzerstorfer
AU  - Robert Tampé
TI  - Dynamic <em>in situ</em> confinement triggers ligand-free neuropeptide receptor signaling
AID  - 10.1101/2021.12.15.472742
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2021.12.15.472742
4099  - http://biorxiv.org/content/early/2022/05/11/2021.12.15.472742.short
4100  - http://biorxiv.org/content/early/2022/05/11/2021.12.15.472742.full
AB  - Membrane receptors are central to cell-cell communication. Receptor clustering at the plasma membrane modulates physiological responses, and mesoscale receptor organization is critical for downstream signaling. Spatially restricted cluster formation of the neuropeptide Y2 hormone receptor (Y2R) was observed in vivo; however, the relevance of this confinement is not fully understood. Here, we controlled Y2R clustering in situ by a chelator nanotool. Due to the multivalent interaction, we observed a dynamic exchange in the microscale confined regions. Fast Y2R enrichment in clustered areas triggered a ligand-independent downstream signaling determined by an increase in cytosolic calcium, cell spreading, and migration. We revealed that the cell response to ligand-induced activation was amplified when cells were pre-clustered by the nanotool. Ligand-independent signaling by clustering differed from ligand-induced activation in the binding of arrestin-3 as downstream effector, which was recruited to the confined regions only in the presence of the ligand. This approach enables in situ clustering of membrane receptors and raises the possibility to explore different modalities of receptor activation.Competing Interest StatementThe authors have declared no competing interest.