Dynamic Submicroscopic Signaling Zones Revealed by Pair Correlation Tracking and Localization Microscopy

Autor(en): You, Changjiang 
Richter, Christian P.
Loechte, Sara
Wilmes, Stephan
Piehler, Jacob 
Stichwörter: BIOTIN LIGASE; CELL-MEMBRANES; Chemistry; Chemistry, Analytical; IMAGE CORRELATION SPECTROSCOPY; LIPID RAFTS; LIVE CELLS; PLASMA-MEMBRANE; PROTEIN HETEROGENEITY; QUANTUM DOTS; SINGLE-MOLECULE TRACKING; SURFACE-PROTEINS
Erscheinungsdatum: 2014
Herausgeber: AMER CHEMICAL SOC
Journal: ANALYTICAL CHEMISTRY
Volumen: 86
Ausgabe: 17
Startseite: 8593
Seitenende: 8602
Zusammenfassung: 
Unraveling the spatiotemporal organization of signaling complexes within the context of plasma membrane nanodomains has remained a highly challenging task. Here, we have applied super-resolution image correlation based on tracking and localization microscopy (TALM) for probing transient confinement as well as ligand binding and intracellular effector recruitment of the type I interferon (IFN) receptor in the plasma membrane of live cells. Ligand and receptor were labeled with monofunctional quantum dots, thus allowing long-term tracking with very high spatial and temporal resolution without an artificial receptor cross-linking at the cell surface. Dual-color TALM was employed for visualizing protein protein interactions involved in IFN signaling at both sides of the plasma membrane with high spatial and temporal resolution. By pair correlation analyses based on time-lapse TALM images (pcTALM), complex assembly within dynamic submicroscopic zones was identified. Strikingly, recruitment of the IFN effector protein signal transducer and activator of transcription 2 (STAT2) into these dynamic signaling zones could be observed. The results suggest that confined diffusion zones in the plasma membrane are employed as transient platforms for the assembly of signaling complexes.
ISSN: 00032700
DOI: 10.1021/ac501127r

Show full item record

Page view(s)

2
Last Week
0
Last month
0
checked on Mar 2, 2024

Google ScholarTM

Check

Altmetric