Long-term single molecule localization microscopy uncovers dynamic co-assembly of Lrp6 and Ror2 into Wnt-signalosomes DOI Creative Commons

Michael Philippi,

Julia Dohle,

Isabelle Watrinet

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 22, 2024

Abstract The conserved Wnt signaling has been classified as two categories of canonical and noncanonical signaling. With a high promiscuity signaling, how receptors from the distinct pathways re-arrange in multi-protein signalosomes remains elusive. We here developed single-molecule tracking localization microscopy based on labeling with reversibly binding nanobodies (rbTALM) for imaging receptor dynamics plasma membrane extended time periods. To this end, we engineered nanobody-tag pairs fine-tuned stabilities ensuring fidelity, yet continuous exchange photobleached labels. Multicolor rbTALM enabled simultaneous super-resolution three different co-receptors same cell more than one hour at video rate. Time-lapse correlation analyses uncovered cooperative association into common, hybrid signalosome, demonstrating exciting possibilities exploring nanoscale across millisecond to timescales.

Language: Английский

Long-term single molecule localization microscopy uncovers dynamic co-assembly of Lrp6 and Ror2 into Wnt-signalosomes DOI Creative Commons

Michael Philippi,

Julia Dohle,

Isabelle Watrinet

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 22, 2024

Abstract The conserved Wnt signaling has been classified as two categories of canonical and noncanonical signaling. With a high promiscuity signaling, how receptors from the distinct pathways re-arrange in multi-protein signalosomes remains elusive. We here developed single-molecule tracking localization microscopy based on labeling with reversibly binding nanobodies (rbTALM) for imaging receptor dynamics plasma membrane extended time periods. To this end, we engineered nanobody-tag pairs fine-tuned stabilities ensuring fidelity, yet continuous exchange photobleached labels. Multicolor rbTALM enabled simultaneous super-resolution three different co-receptors same cell more than one hour at video rate. Time-lapse correlation analyses uncovered cooperative association into common, hybrid signalosome, demonstrating exciting possibilities exploring nanoscale across millisecond to timescales.

Language: Английский

Citations

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