Global phosphoproteomic effects of natural tyrosine kinase inhibitor, genistein, on signaling pathways.
Genistein is a natural protein tyrosine kinase inhibitor that exerts anti-cancer effect by inducing G2/M arrest and apoptosis. However, the phosphotyrosine signaling pathways mediated by genistein are largely unknown. In this study, we combined tyrosine phosphoprotein enrichment with MS-based quantitative proteomics technology to globally identify genistein-regulated tyrosine phosphoproteins aiming to depict genistein-inhibited phosphotyrosine cascades. Our experiments resulted in the identification of 213 phosphotyrosine sites on 181 genistein-regulated proteins. Many identified phosphoproteins, including nine protein kinases, eight receptors, five protein phosphatases, seven transcriptical regulators and four signal adaptors, were novel inhibitory effectors with no previously known function in the anti-cancer mechanism of genistein. Functional analysis suggested that genistein-regulated protein tyrosine phosphorylation mainly by inhibiting the activity of tyrosine kinase EGFR, PDGFR, insulin receptor, Abl, Fgr, Itk, Fyn and Src. Core signaling molecules inhibited by genistein can be functionally categorized into the canonial Receptor-MAPK or Receptor-PI3K/AKT cascades. The method used here may be suitable for the identification of inhibitory effectors and tyrosine kinases regulated by anti-cancer drugs.
Yan
Guang-Rong
GR
Institute of Life and Health Engineering, and National Engineering and Research Center for Genetic Medicine, Jinan University, Guangzhou, P R China.
Xiao
Chuan-Le
CL
He
Gui-Wei
GW
Yin
Xing-Feng
XF
Chen
Nan-Peng
NP
Cao
Ya
Y
He
Qing-Yu
QY
D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000187:drug effects; D016195:G2 Phase / Q000187:drug effects; D019833:Genistein / Q000494:pharmacology*; D006801:Humans; D007553:Isotope Labeling; D013058:Mass Spectrometry; D008938:Mitosis / Q000187:drug effects; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects*
D010455:Peptides; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D019000:Phosphotyrosine; D019833:Genistein; D011505:Protein-Tyrosine Kinases