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  Heterogeneity of signal transduction at the subcellular level: microsphere-based focal EGF receptor activation and stimulation of Shc translocation

Brock, R., & Jovin, T. M. (2001). Heterogeneity of signal transduction at the subcellular level: microsphere-based focal EGF receptor activation and stimulation of Shc translocation. Journal of Cell Science, 114, 2437-2447.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F5AE-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-9E35-0
Genre: Journal Article

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 Creators:
Brock, R.1, Author              
Jovin, T. M.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: green fluorescent protein; GFP; Venus; fluorescence polarization
 Abstract: Epidermal growth factor receptor (EGFR, erbB1) activation and translocation of the Shc adaptor protein to activated receptors were analyzed at the subcellular level by dual-label immunofluorescence and confocal laser scanning microscopy in conjunction with a new microsphere-based protocol. In the Quantitative Microsphere Recruitment Assay (QMRA) introduced here, epidermal growth factor-coated 1 mm diameter microspheres were distributed over the surface of adherent tissue culture cells expressing the receptor. High-resolution confocal microscopy of a fusion construct of the receptor and the green fluorescent protein expressed in Chinese hamster ovary cells demonstrated that engulfment and internalization of the microspheres occurred rapidly within minutes, and in a receptor activation-dependent manner. In human epidermoid carcinoma A431 cells, receptor activation and Shc translocation persisted over the 20-minute time course of the experiments. However, at the subcellular level the positive correlation of receptor activation and Shc translocation observed at 5-8 minutes dissipated, indicating a time-dependent decoupling of thetwo events and variation in the kinetics of signal transduction for different subcellular locations.

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Language(s): eng - English
 Dates: 2005-08-182001
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 223393
Other: 34671
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Title: Journal of Cell Science
Source Genre: Journal
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Pages: - Volume / Issue: 114 Sequence Number: - Start / End Page: 2437 - 2447 Identifier: -