Comparisons of tyrosine phosphorylated proteins in cells expressing lung cancer-specific alleles of EGFR and KRAS Journal Article
| Authors: | Guha, U.; Chaerkady, R.; Marimuthu, A.; Patterson, A. S.; Kashyap, M. K.; Harsha, H. C.; Sato, M.; Bader, J. S.; Lash, A. E.; Minna, J. D.; Pandey, A.; Varmus, H. E. |
| Article Title: | Comparisons of tyrosine phosphorylated proteins in cells expressing lung cancer-specific alleles of EGFR and KRAS |
| Abstract: | We have used unbiased phosphoproteomic approaches, based on quantitative mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC), to identify tyrosine phosphorylated proteins in isogenic human bronchial epithelial cells (HBECs) and human lung adenocarcinoma cell lines, expressing either of the two mutant alleles of EGFR (L858R and Del E746-A750), or a mutant KRAS allele, which are common in human lung adenocarcinomas. Tyrosine phosphorylation of signaling molecules was greater in HBECs expressing the mutant EGFRs than in cells expressing WT EGFR or mutant KRAS. Receptor tyrosine kinases (such as EGFR, ERBB2, MET, and IGF1R), and Mig-6, an inhibitor of EGFR signaling, were more phosphorylated in HBECs expressing mutant EGFR than in cells expressing WT EGFR or mutant RAS. Phosphorylation of some proteins differed in the two EGFR mutant-expressing cells; for example, some cell junction proteins (β-catenin, plakoglobin, and E-cadherin) were more phosphorylated in HBECs expressing L858R EGFR than in cells expressing Del EGFR. There were also differences in degree of phosphorylation at individual tyrosine sites within a protein; for example, a previously uncharacterized phosphorylation site in the nucleotide-binding loop of the kinase domains of EGFR (Y727), ERBB2 (Y735), or ERBB4 (Y733), is phosphorylated significantly more in HBECs expressing the deletion mutant than in cells expressing the wild type or L858R EGFR. Signaling molecules not previously implicated in ERBB signaling, such as polymerase transcript release factor (PTRF), were also phosphorylated in cells expressing mutant EGFR. Bayesian network analysis of these and other datasets revealed that PTRF might be a potentially important component of the ERBB signaling network. © 2008 by The National Academy of Sciences of the USA. |
| Keywords: | signal transduction; protein expression; protein phosphorylation; human cell; mutation; proto-oncogene proteins; mass spectrometry; allele; cells, cultured; bayes theorem; lung neoplasms; epidermal growth factor receptor; epidermal growth factor receptor 2; protein; protein binding; alleles; lung cancer; receptor, epidermal growth factor; cancer cell culture; protein tyrosine kinase; tyrosine; proteomics; uvomorulin; endothelial cells; gene expression regulation, neoplastic; amino acid sequence; isotope labeling; cell culture; quantitative analysis; immunoprecipitation; tandem mass spectrometry; high performance liquid chromatography; ras proteins; cell junction; k ras protein; beta catenin; bronchi; cell mutant; cell labeling; epidermal growth factor receptor 4; phosphotyrosine; cell immortalization; tyrosine phosphorylation; nucleotide binding site; lung alveolus cell; plakoglobin; comparative anatomy |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 105 |
| Issue: | 37 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2008-09-16 |
| Start Page: | 14112 |
| End Page: | 14117 |
| Language: | English |
| DOI: | 10.1073/pnas.0806158105 |
| PUBMED: | 18776048 |
| PROVIDER: | scopus |
| PMCID: | PMC2531065 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 27" - "Export Date: 17 November 2011" - "CODEN: PNASA" - "Source: Scopus" |
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