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

Crystal L. Woodard, C. Rory Goodwin, Jun Wan, Shuli Xia, Robert Newman, Jianfei Hu, Jin Zhang, S. Diane Hayward, Jiang Qian, John J Laterra, Heng Zhu

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article numbere72671
JournalPLoS One
Volume8
Issue number9
StatePublished - 2013
Externally publishedYes

Fingerprint

hepatocyte growth factor
Hepatocyte Growth Factor
protein phosphorylation
Phosphorylation
Protein Array Analysis
Microarrays
Proteins
proteins
Tissue
phosphorylation
cells
Proto-Oncogene Proteins c-met
proteome
carcinogenesis
Chemical activation
Proteome
morphogenesis
tyrosine
Oligonucleotide Array Sequence Analysis
Morphogenesis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Woodard, C. L., Goodwin, C. R., Wan, J., Xia, S., Newman, R., Hu, J., ... Zhu, H. (2013). Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling. PLoS One, 8(9), [e72671].

Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling. / Woodard, Crystal L.; Goodwin, C. Rory; Wan, Jun; Xia, Shuli; Newman, Robert; Hu, Jianfei; Zhang, Jin; Hayward, S. Diane; Qian, Jiang; Laterra, John J; Zhu, Heng.

In: PLoS One, Vol. 8, No. 9, e72671, 2013.

Research output: Contribution to journalArticle

Woodard, CL, Goodwin, CR, Wan, J, Xia, S, Newman, R, Hu, J, Zhang, J, Hayward, SD, Qian, J, Laterra, JJ & Zhu, H 2013, 'Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling.', PLoS One, vol. 8, no. 9, e72671.
Woodard CL, Goodwin CR, Wan J, Xia S, Newman R, Hu J et al. Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling. PLoS One. 2013;8(9). e72671.
Woodard, Crystal L. ; Goodwin, C. Rory ; Wan, Jun ; Xia, Shuli ; Newman, Robert ; Hu, Jianfei ; Zhang, Jin ; Hayward, S. Diane ; Qian, Jiang ; Laterra, John J ; Zhu, Heng. / Profiling the dynamics of a human phosphorylome reveals new components in HGF/c-Met signaling. In: PLoS One. 2013 ; Vol. 8, No. 9.
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