Probing the Heterogeneity of Protein Kinase Activation in Cells by Super-resolution Microscopy

Title: Probing the Heterogeneity of Protein Kinase Activation in Cells by Super-resolution Microscopy
Authors: Zhang, RuobingFruhwirth, Gilbert O.Coban, OanaKholodenko, Boris al.
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Date: 21-Oct-2016
Online since: 2019-04-01T10:43:07Z
Abstract: Heterogeneity of mitogen-activated protein kinase (MAPK) activation in genetically identical cells, which occurs in response to epidermal growth factor receptor (EGFR) signaling, remains poorly understood. MAPK cascades integrate signals emanating from different EGFR spatial locations, including the plasma membrane and endocytic compartment. We previously hypothesized that in EGF-stimulated cells the MAPK phosphorylation (pMAPK) level and activity are largely determined by the spatial organization of the EGFR clusters within the cell. For experimental testing of this hypothesis, we used super-resolution microscopy to define EGFR clusters by receptor numbers (N) and average intracluster distances (d). From these data, we predicted the extent of pMAPK with 85% accuracy on a cell-to-cell basis with control data returning 54% accuracy (P < 0.001). For comparison, the prediction accuracy was only 61% (P = 0.382) when the diffraction-limited averaged fluorescence intensity/cluster was used. Large clusters (N ≥ 3) with d > 50 nm were most predictive for pMAPK level in cells. Electron microscopy revealed that these large clusters were primarily localized to the limiting membrane of multivesicular bodies (MVB). Many tighter packed dimers/multimers (d < 50 nm) were found on intraluminal vesicles within MVBs, where they were unlikely to activate MAPK because of the physical separation. Our results suggest that cell-to-cell differences in N and d contain crucial information to predict EGFR-activated cellular pMAPK levels and explain pMAPK heterogeneity in isogenic cells.
Funding Details: European Commission - Seventh Framework Programme (FP7)
Wellcome Trust
Funding Details: National Institutes of Health
Dimbleby Cancer Care
KCL-UCL Comprehensive Cancer Imaging Centre (CCIC)
Type of material: Journal Article
Publisher: American Chemical Society
Journal: ACS Nano
Volume: 11
Issue: 1
Start page: 249
End page: 257
Copyright (published version): 2016 the American Chemical Society
Keywords: Bayesian modelingCell-to-cell heterogeneityEGFRMAPKSuper-resolution microscopy
DOI: 10.1021/acsnano.6b05356
Language: en
Status of Item: Peer reviewed
This item is made available under a Creative Commons License:
Appears in Collections:Conway Institute Research Collection
SBI Research Collection
Medicine Research Collection

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