%0 Journal Article %1 lill2005kinetics %A Lill, Yoriko %A Martinez, Karen L. %A Lill, Markus A. %A Meyer, Bruno H. %A Vogel, Horst %A Hecht, Bert %D 2005 %J ChemPhysChem %K experiment nano-optics single-molecules %N 8 %P 1633-1640 %R 10.1002/cphc.200500111 %T Kinetics of the Initial Steps of G Protein-Coupled Receptor-Mediated Cellular Signaling Revealed by Single-Molecule Imaging %V 6 %X We report on an in vivo single-molecule study of the signaling kinetics of G protein-coupled receptors (GPCR) performed using the neurokinin 1 receptor (NK1R) as a representative member. The NK1R signaling cascade is triggered by the specific binding of a fluorescently labeled agonist, substance P (SP). The diffusion of single receptor–ligand complexes in plasma membrane of living HEK 293 cells is imaged using fast single-molecule wide-field fluorescence microscopy at 100 ms time resolution. Diffusion trajectories are obtained which show intra- and intertrace heterogeneity in the diffusion mode. To investigate universal patterns in the diffusion trajectories we take the ligand-binding event as the common starting point. This synchronization allows us to observe changes in the character of the ligand–receptor-complex diffusion. Specifically, we find that the diffusion of ligand–receptor complexes is slowed down significantly and becomes more constrained as a function of time during the first 1000 ms. The decelerated and more constrained diffusion is attributed to an increasing interaction of the GPCR with cellular structures after the ligand–receptor complex is formed.

@article{lill2005kinetics, abstract = {We report on an in vivo single-molecule study of the signaling kinetics of G protein-coupled receptors (GPCR) performed using the neurokinin 1 receptor (NK1R) as a representative member. The NK1R signaling cascade is triggered by the specific binding of a fluorescently labeled agonist, substance P (SP). The diffusion of single receptor–ligand complexes in plasma membrane of living HEK 293 cells is imaged using fast single-molecule wide-field fluorescence microscopy at 100 ms time resolution. Diffusion trajectories are obtained which show intra- and intertrace heterogeneity in the diffusion mode. To investigate universal patterns in the diffusion trajectories we take the ligand-binding event as the common starting point. This synchronization allows us to observe changes in the character of the ligand–receptor-complex diffusion. Specifically, we find that the diffusion of ligand–receptor complexes is slowed down significantly and becomes more constrained as a function of time during the first 1000 ms. The decelerated and more constrained diffusion is attributed to an increasing interaction of the GPCR with cellular structures after the ligand–receptor complex is formed.}, added-at = {2020-02-19T12:22:33.000+0100}, author = {Lill, Yoriko and Martinez, Karen L. and Lill, Markus A. and Meyer, Bruno H. and Vogel, Horst and Hecht, Bert}, biburl = {https://www.bibsonomy.org/bibtex/2e6724bb2ad28013e47813ad086eda52f/ep5optics}, copyright = {Copyright © 2005 WILEY‐VCH Verlag GmbH \& Co. KGaA, Weinheim}, day = 05, doi = {10.1002/cphc.200500111}, file = {Lill et al. - 2005 - Kinetics of the Initial Steps of G Protein-Coupled.pdf:C\:\\Users\\scherzad\\Zotero\\storage\\JLTW24HI\\Lill et al. - 2005 - Kinetics of the Initial Steps of G Protein-Coupled.pdf:application/pdf;Snapshot:C\:\\Users\\scherzad\\Zotero\\storage\\LGPTUP4X\\cphc.html:text/html}, interhash = {19646e498c8768d487b5b6465908a538}, intrahash = {e6724bb2ad28013e47813ad086eda52f}, issn = {1439-7641}, journal = {ChemPhysChem}, keywords = {experiment nano-optics single-molecules}, language = {en}, month = {08}, number = 8, pages = {1633-1640}, timestamp = {2020-02-19T12:22:33.000+0100}, title = {Kinetics of the Initial Steps of G Protein-Coupled Receptor-Mediated Cellular Signaling Revealed by Single-Molecule Imaging}, urldate = {2020-02-19}, volume = 6, year = 2005 }