Journal
NATURE METHODS
Volume 7, Issue 2, Pages 148-U95Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1418
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Funding
- University of Chicago Cancer Research Center
- American Cancer Society
- University of Chicago Breast Cancer
- Cancer Research Foundation
- Illinois Department of Health [86280156]
- US National Institutes of General Medical Sciences [P50-GM0686762]
- US National Cancer Institute [CA96504]
- National Institutes of Health
- National Science Foundation
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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.
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