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

Zhu, Heng

Koul, Hari

United States

2017-12-02T17:22:27Z

2017-12-02T17:22:27Z

2013

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.

https://www.ncbi.nlm.nih.gov/pubmed/24023761

PONE-D-12-27318

1932-6203

https://hdl.handle.net/10161/15823

eng

Public Library of Science (PLoS)

PLoS One

10.1371/journal.pone.0072671

Hepatocyte Growth Factor

Humans

Oligonucleotide Array Sequence Analysis

Phosphorylation

Protein Array Analysis

Proto-Oncogene Proteins c-met

Signal Transduction

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

Journal article

Goodwin, C Rory|0000-0002-6540-2751

https://www.ncbi.nlm.nih.gov/pubmed/24023761

e72671

9

Clinical Science Departments

Duke

Neurosurgery

School of Medicine

Temp group - logins allowed

Published online

8