Basic study on SH2 domain of Grb2 as a molecular probe for detection of RTK activation

  • Authors: Yuriko Saito, Takako Furukawa, Yasushi Arano, Yasuhisa Fujibayashi, Tsuneo Saga
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  • Published online on: Sunday, August 1, 2010
  • Pages: 281-287
  • DOI: 10.3892/ijo_00000676

Abstract

Epidermal growth factor receptor (EGFR) and other receptor tyrosine kinases (RTKs) are overexpressed and/or mutated in various cancers and their abnormal activation is implicated in carcinogenesis. We explored the possibility of generating an imaging probe for RTK activation using the SH2 domain of Grb2 for cancer characterization. For cell penetration, molecular analysis and radiolabeling, the SH2 domain was fused with TAT, flag and tyrosine residue, respectively (termed TSF). We analyzed TSF characteristics in cells such as cellular uptake, stability and localization. After uptake into EGFR-expressing cells, TSF was found to be binding to phosphorylated-EGFR, which increased by stimulation with EGF. TSF was co-localized with EGFR in EGFR-activated cells, while it was localized as dots in cytosol in EGFR-non-activated cells. Cellular retention time of TSF was significantly extended under EGFR activation with EGF stimuli and reduced under the treatment with a tyrosine kinase inhibitor, Tyrphostin AG1478. In conclusion, the SH2 domain of Grb2 has the potential to be used as a binding component of a probe to detect activated-RTK and to evaluate the effect of kinase inhibitors on RTK activation.
Journal Cover

August 2010
Volume 37 Issue 2

Print ISSN: 1019-6439
Online ISSN:1791-2423

2012 Impact Factor: 2.773
Ranked #30/202 Oncology
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APA
Saito, Y., Furukawa, T., Arano, Y., Fujibayashi, Y., & Saga, T. (2010). Basic study on SH2 domain of Grb2 as a molecular probe for detection of RTK activation. International Journal of Oncology, 37(2), 281-287.
MLA
Saito, Furukawa, Arano, Fujibayashi, and Tsuneo Saga. "Basic study on SH2 domain of Grb2 as a molecular probe for detection of RTK activation." International Journal of Oncology International Journal of Oncology 37.2 (2010): 281-287.
Chicago
Saito, Furukawa, Arano, Fujibayashi, and Tsuneo Saga. "Basic study on SH2 domain of Grb2 as a molecular probe for detection of RTK activation." International Journal of Oncology International Journal of Oncology 37 no. 2 (2010): 281-287.