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Proteomic analysis of novel targets associated with TrkA‐mediated tyrosine phosphorylation signaling pathways in SK ‐ N ‐ MC neuroblastoma cells
Author(s) -
Jung Eun Joo,
Lee SangYeul,
Kim Choong Won
Publication year - 2013
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201200251
Subject(s) - tropomyosin receptor kinase a , phosphorylation , low affinity nerve growth factor receptor , microbiology and biotechnology , tyrosine phosphorylation , receptor tyrosine kinase , biology , signal transduction , trk receptor , tyrosine kinase , tyrosine , chemistry , nerve growth factor , biochemistry , receptor
Tropomyosin‐related kinase A ( T rk A ) is a receptor‐type protein tyrosine kinase and exploits pleiotypic roles via nerve growth factor (NGF)‐dependent or NGF‐independent mechanisms in various cell types. Here, we showed that the inhibition of T rk A activity by GW 441756 resulted in the suppression of tyrosine phosphorylation of cellular proteins including extracellular signal‐regulated protein kinase (ERK) and c‐ J un N ‐terminal kinase (JNK). To find novel targets associated with T rk A ‐mediated tyrosine phosphorylation signaling pathways, we investigated GW 441756 effects on T rk A ‐dependent targets in SK ‐ N ‐ MC neuroblastoma cells by proteomic analysis. The major T rk A ‐dependent protein spots controlled by GW 441756 were determined by PDQ uest image analysis, identified by MALDI ‐ TOF MS and MALDI ‐ TOF / TOF MS / MS , and verified by 2 DE / W estern blot analysis. Thus, we found that most of the identified protein spots were modified forms in a normal condition, and their modifications were regulated by T rk A activity. Especially, our results demonstrated that the modifications of α‐tubulin and heterogeneous nuclear ribonucleoproteins C 1/ C 2 (hnRNP C1/C2) were significantly upregulated by T rk A , whereas α‐enolase modification was downregulated by T rk A , and it was suppressed by GW 441756, indicating that T rk A activity is required for their modifications. Taken together, we suggest here that the major novel T rk A ‐dependent targets such as α‐tubulin, hnRNP C1/C2, and α‐enolase could play an essential role in T rk A ‐mediated tyrosine phosphorylation signaling pathways via regulation of their posttranslational modifications.