Identification of tyrosine-phosphorylated proteins associated with lung cancer metastasis using label-free quantitative analyses.
Lung cancer is a lethal disease, and early metastasis is the major cause of treatment failure and cancer-related death. Tyrosine phosphorylated (P-Tyr) proteins are involved in the invasive and metastatic behavior of lung cancer; however, only a limited number of targets were identified. We attempt to characterize P-Tyr proteins and events involved in the metastatic process. In a previous work, we have developed a strategy for identification of protein phosphorylation. Here, this strategy was used to characterize the tyrosine phosphoproteome of lung cancer cells that have different invasive abilities (CL1-0 vs. CL1-5). Using our analytical strategy, we report the identification of 335 P-Tyr sites from 276 phosphoproteins. Label-free quantitative analysis revealed that 36 P-Tyr peptides showed altered levels between CL1-0 and CL1-5 cells. From this list of sites, we extracted two novel consensus sequences and four known motifs for specific kinases and phosphatases including EGFR, Src, JAK2, and TC-PTP. Protein-protein interaction network analysis of the altered P-Tyr proteins illustrated that 11 proteins were linked to a network containing EGFR, c-Src, c-Myc, and STAT, which is known to be related to lung cancer metastasis. Among these 11 proteins, 7 P-Tyr proteins have not been previously reported to be associated with lung cancer metastasis and are of greatest interest for further study. The characterized tyrosine phosphoproteome and altered P-Tyr targets may provide a better comprehensive understanding of the mechanisms of lung cancer invasion/metastasis and discover potential therapies.
Wu
Hsin-Yi
HY
Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
Tseng
Vincent S
VS
Chen
Lien-Chin
LC
Chang
Hui-Yin
HY
Chuang
I-Chi
IC
Tsay
Yeou-Guang
YG
Liao
Pao-Chi
PC
D000469:Alkaline Phosphatase; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D019295:Computational Biology; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D053614:Janus Kinase 2 / Q000378:metabolism; D008175:Lung Neoplasms / Q000473:pathology*; D009362:Neoplasm Metastasis / Q000175:diagnosis*; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism*; D010766:Phosphorylation; D054578:Protein Tyrosine Phosphatase, Non-Receptor Type 2 / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D014025:Titanium; D014443:Tyrosine / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism
D010750:Phosphoproteins; C009495:titanium dioxide; D014443:Tyrosine; D014025:Titanium; C512478:EGFR protein, human; D066246:ErbB Receptors; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D019061:src-Family Kinases; D000469:Alkaline Phosphatase; C516649:PTPN2 protein, human; D054578:Protein Tyrosine Phosphatase, Non-Receptor Type 2