Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling.
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 articleSubject
Hepatocyte Growth FactorHumans
Oligonucleotide Array Sequence Analysis
Phosphorylation
Protein Array Analysis
Proto-Oncogene Proteins c-met
Signal Transduction
Permalink
https://hdl.handle.net/10161/15823Published Version (Please cite this version)
10.1371/journal.pone.0072671Publication 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.
Collections
More Info
Show full item recordScholars@Duke
Courtney Rory Goodwin
Assistant Professor of Neurosurgery

Articles written by Duke faculty are made available through the campus open access policy. For more information see: Duke Open Access Policy
Rights for Collection: Scholarly Articles