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Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling.

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Date
2013
Authors
Woodard, Crystal L
Goodwin, C Rory
Wan, Jun
Xia, Shuli
Newman, Robert
Hu, Jianfei
Zhang, Jin
Hayward, S Diane
Qian, Jiang
Laterra, John
Zhu, Heng
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Abstract
Protein phosphorylation is a dynamic and reversible event that greatly influences cellular function. Identifying the key regulatory elements that determine cellular phenotypes during development and oncogenesis requires the ability to dynamically monitor proteome-wide events. Here, we report the development of a new strategy to monitor dynamic changes of protein phosphorylation in cells and tissues using functional protein microarrays as the readout. To demonstrate this technology's ability to identify condition-dependent phosphorylation events, human protein microarrays were incubated with lysates from cells or tissues under activation or inhibition of c-Met, a receptor tyrosine kinase involved in tissue morphogenesis and malignancy. By comparing the differences between the protein phosphorylation profiles obtained using the protein microarrays, we were able to recover many of the proteins that are known to be specifically activated (i.e., phosphorylated) upon c-Met activation by the hepatocyte growth factor (HGF). Most importantly, we discovered many proteins that were differentially phosphorylated by lysates from cells or tissues when the c-Met pathway was active. Using phosphorylation-specific antibodies, we were able to validate several candidate proteins as new downstream components of the c-Met signaling pathway in cells. We envision that this new approach, like its DNA microarray counterpart, can be further extended toward profiling dynamics of global protein phosphorylation under many different physiological conditions both in cellulo and in vivo in a high-throughput and cost-effective fashion.
Type
Journal article
Subject
Hepatocyte Growth Factor
Humans
Oligonucleotide Array Sequence Analysis
Phosphorylation
Protein Array Analysis
Proto-Oncogene Proteins c-met
Signal Transduction
Permalink
https://hdl.handle.net/10161/15823
Published Version (Please cite this version)
10.1371/journal.pone.0072671
Publication Info
Woodard, Crystal L; Goodwin, C Rory; Wan, Jun; Xia, Shuli; Newman, Robert; Hu, Jianfei; ... Zhu, Heng (2013). Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling. PLoS One, 8(9). pp. e72671. 10.1371/journal.pone.0072671. Retrieved from https://hdl.handle.net/10161/15823.
This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.
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Scholars@Duke

Goodwin

Courtney Rory Goodwin

Assistant Professor of Neurosurgery
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