Actinomycin D as a novel SH2 domain ligand inhibits Shc/Grb2 interaction in B104‐1‐1 (neu*‐transformed NIH3T3) and SAA (hEGFR‐overexpressed NIH3T3) cells

FEBS Letters

Bok, Song Hae²  Han, Mi Young²  Kwon, Byoung-Mog²  Lee, Eun Kyung²  Choi, Jung-Do¹  Kim, Hyae-Kyeong²  Nam, Ji-Youn² 

[1] Department of Biochemistry, Chungbuk National University, Cheongju 361-763, South Korea;Korea Research Institute of Bioscience and Biotechnology, KIST, P.O. Box 115, Yusong, Taejon 305-600, South Korea

关键词: Grb2;    Shc;    Actinomycin;    Src homology 2 domain;    Cyclopeptide antibiotic;    Extracellular signal-regulated protein kinase;    SH2;    src homology 2;    MAP kinase;    mitogen-activated protein kinase;    Erk;    extracellular signal-regulated protein kinase;    hEGFR;    human epidermal growth factor receptor;    FBS;    fetal bovine serum;   

DOI  :  10.1016/S0014-5793(99)00710-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Actinomycins, a family of bicyclic chromopeptide lactones with strong antineoplastic activity, were screened as inhibitors of Shc/Grb2 interaction in in vitro assay systems. To investigate the effects of actinomycin D on Shc/Grb2 interaction in cell-based experiments, we used SAA (normal hEGFR-overexpressed NIH3T3) cells and B104-1-1 (neu*-transformed NIH3T3) cells, because a large number of the Shc/Grb2 complexes were detected. Associated protein complexes containing Shc were immunoprecipitated from actinomycin D-treated cell lysates with polyclonal anti-Shc antibody. Then the association with Grb2 was assessed by immunoblotting with monoclonal anti-Grb2 antibody. The result of the immunoblotting experiment revealed that actinomycin D inhibited Shc/Grb2 interaction in a dose-dependent manner in both B104-1-1 and EGF-stimulated SAA cells. The inhibition of Shc/Grb2 interaction by actinomycin D in B104-1-1 cells also reduced tyrosine phosphorylation of MAP kinase (Erk1/Erk2), one of the major components in the Ras-MAP kinase signaling pathway. These results suggest that actinomycin D could be a non-phosphorylated natural and cellular membrane-permeable SH2 domain antagonist.

【 授权许可】

Unknown