Systems analysis of EGF receptor signaling dynamics with microwestern arrays.

Authors: Ciaccio MF; Wagner JP; Chuu CP; Lauffenburger DA; Jones RB

Abstract: We describe microwestern arrays, which enable quantitative, sensitive and high-throughput assessment of protein abundance and modifications after electrophoretic separation of microarrayed cell lysates. This method allowed us to measure 91 phosphosites on 67 proteins at six time points after stimulation with five epidermal growth factor (EGF) concentrations in A431 human carcinoma cells. We inferred the connectivities among 15 phosphorylation sites in 10 receptor tyrosine kinases (RTKs) and two sites from Src kinase using Bayesian network modeling and two mutual information-based methods; the three inference methods yielded substantial agreement on the network topology. These results imply multiple distinct RTK coactivation mechanisms and support the notion that small amounts of experimental data collected from phenotypically diverse network states may enable network inference.

Keywords: Blotting, Western/*instrumentation/methods; Equipment Design; Equipment Failure Analysis; Protein Array Analysis/*instrumentation/methods; Proteome/*metabolism; Receptor, Epidermal Growth Factor/*metabolism; Reproducibility of Results; Sensitivity and Specificity; Signal Transduction/*physiology; Systems Biology
Journal: Nature methods
Volume: 7
Issue: 2
Pages: 148-55
Date: Jan. 27, 2010
PMID: 20101245
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Citation:

Ciaccio MF, Wagner JP, Chuu CP, Lauffenburger DA, Jones RB (2010) Systems analysis of EGF receptor signaling dynamics with microwestern arrays. Nature methods 7: 148-55.



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