Detail of Fig4A_EGFR-GFP_GRB2



Project
Title
Time-lapse of single-molecule imaging of EGFinduced GRB2–HaloTag::TMR translocation from cytoplasm to the basal plasma membrane.
Description
Time-lapse of single-molecule imaging of EGFinduced GRB2–HaloTag::TMR translocation from cytoplasm to the basal plasma membrane.
Release, Updated
2022-03-31
License
CC BY
Kind
Image data
File Formats
.tif
Data size
3.5 GB

Organism
Cricetulus griseus ( NCBI:txid10029 )
Strain(s)
-
Cell Line
CHO-K1 ( CLO_0002462 )
Protein names
GRB2

Datatype
-
Molecular Function (MF)
epidermal growth factor-activated receptor activity ( GO:0005006 ) epidermal growth factor receptor binding ( GO:0005154 )
Biological Process (BP)
epidermal growth factor receptor signaling pathway ( GO:0007173 )
Cellular Component (CC)
Grb2-EGFR complex ( GO:0070436 )
Biological Imaging Method
time lapse microscopy ( Fbbi:00000249 )
X scale
44 nanometer/pixel
Y scale
44 nanometer/pixel
Z scale
-
T scale
30.5 microsecond per time interval

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Hiroshima M, et. al. (2018) J Mol Biol., 430(9):1386-1401.
Related paper(s)

Michio Hiroshima, Chan-Gi Pack, Kazunari Kaizu, Koichi Takahashi, Masahiro Ueda, Yasushi Sako (2018) Transient Acceleration of Epidermal Growth Factor Receptor Dynamics Produces Higher-Order Signaling Clusters., Journal of molecular biology, Volume 430, Number 9, pp. 1386-1401

Published in 2018 Apr 27 (Electronic publication in March 2, 2018, midnight )

(Abstract) Cell signaling depends on spatiotemporally regulated molecular interactions. Although the movements of signaling proteins have been analyzed with various technologies, how spatial dynamics influence the molecular interactions that transduce signals is unclear. Here, we developed a single-molecule method to analyze the spatiotemporal coupling between motility, clustering, and signaling. The analysis was performed with the epidermal growth factor receptor (EGFR), which triggers signaling through its dimerization and phosphorylation after association with EGF. Our results show that the few EGFRs isolated in membrane subdomains were released by an EGF-dependent increase in their diffusion area, facilitating molecular associations and producing immobile clusters. Using a two-color single-molecule analysis, we found that the EGF-induced state transition alters the properties of the immobile clusters, allowing them to interact for extended periods with the cytoplasmic protein, GRB2. Our study reveals a novel correlation between this molecular interaction and its mesoscale dynamics, providing the initial signaling node.
(MeSH Terms)

Contact
Yasushi Sako , RIKEN , Cellular Informatics Laboratory
Contributors
Michio Hiroshima

OMERO Dataset
OMERO Project
Source