FRET analysis of protein tyrosine kinase c-Src activation mediated via aryl hydrocarbon receptor. 2011

Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
Center for Health and the Environment, University of California School of Medicine, USA.

BACKGROUND Activation of the protein tyrosine kinase c-Src (c-Src kinase) induced by the exposure to the environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown in various cell types. Most previous works used Western blot analysis to detect the phosphorylation on the Tyr416 residue, which activates c-Src kinase. METHODS Here we compared the results of c-Src tyrosine phosphorylation via aryl hydrocarbon receptor (AhR)-dependent mechanisms from Western blot analysis with fluorescent resonance energy transfer (FRET) assay detecting c-Src activation after treatment with TCDD to activate AhR in two different human cell types. RESULTS Western blot analyses show time-dependent phosphorylation of c-Src by TCDD in HepG2 and MCF-10A cells. Data from FRET assay visualized and quantified the activation of c-Src kinase induced by TCDD in living cells of both cell types. The FRET efficiency decreased by 20%, 5 min after TCDD treatment and continued decreasing until the end of the experiment, 25 min after TCDD treatment. PP2, a c-Src specific inhibitor, suppressed both TCDD- and epidermal growth factor- (EGF) induced c-Src activation. In contrast, the AhR antagonist 3'-methoxy-4'nitroflavone (MNF) blocked only TCDD- but not EGF-induced activation of c-Src. CONCLUSIONS The current study shows that the early activation of c-Src via EGF and AhR signaling pathways can be visualized in living cells using the FRET assay which is in line with Western blot analysis. CONCLUSIONS The FRET assay provides a useful tool to visualize and quantify c-Src kinase activation via AhR in living cells.

UI MeSH Term Description Entries
D010766 Phosphorylation The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety. Phosphorylations
D011505 Protein-Tyrosine Kinases Protein kinases that catalyze the PHOSPHORYLATION of TYROSINE residues in proteins with ATP or other nucleotides as phosphate donors. Tyrosine Protein Kinase,Tyrosine-Specific Protein Kinase,Protein-Tyrosine Kinase,Tyrosine Kinase,Tyrosine Protein Kinases,Tyrosine-Specific Protein Kinases,Tyrosylprotein Kinase,Kinase, Protein-Tyrosine,Kinase, Tyrosine,Kinase, Tyrosine Protein,Kinase, Tyrosine-Specific Protein,Kinase, Tyrosylprotein,Kinases, Protein-Tyrosine,Kinases, Tyrosine Protein,Kinases, Tyrosine-Specific Protein,Protein Kinase, Tyrosine-Specific,Protein Kinases, Tyrosine,Protein Kinases, Tyrosine-Specific,Protein Tyrosine Kinase,Protein Tyrosine Kinases,Tyrosine Specific Protein Kinase,Tyrosine Specific Protein Kinases
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D004785 Environmental Pollutants Substances or energies, for example heat or light, which when introduced into the air, water, or land threaten life or health of individuals or ECOSYSTEMS. Environmental Pollutant,Pollutant,Pollutants,Pollutants, Environmental,Pollutant, Environmental
D004789 Enzyme Activation Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme. Activation, Enzyme,Activations, Enzyme,Enzyme Activations
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000072317 Polychlorinated Dibenzodioxins Dibenzodioxin derivatives that contain multiple chloride atoms bound to the benzene ring structures. TCDD,Tetrachlorodibenzodioxin,2,3,7,8-Tetrachlorodibenzo-p-dioxin,Chlorinated Dibenzo-p-dioxins,Dibenzo(b,e)(1,4)dioxin, 2,3,7,8-tetrachloro-,PCDD,Polychlorinated Dibenzo-p-dioxins,Polychlorinated Dibenzodioxin,Polychlorodibenzo-4-dioxin,Polychlorodibenzo-p-dioxin,Tetrachlorodibenzo-p-dioxin,Chlorinated Dibenzo p dioxins,Dibenzo-p-dioxins, Chlorinated,Dibenzo-p-dioxins, Polychlorinated,Dibenzodioxin, Polychlorinated,Dibenzodioxins, Polychlorinated,Polychlorinated Dibenzo p dioxins,Polychlorodibenzo 4 dioxin,Polychlorodibenzo p dioxin,Tetrachlorodibenzo p dioxin
D000081247 CSK Tyrosine-Protein Kinase Protein tyrosine kinases that phosphorylate tyrosine residues located in the C-terminal tails of SRC-FAMILY KINASES. C-Terminal Src Kinase,Carboxy-Terminal Src Kinase,Protein Tyrosine Kinase p50(csk),Protein-Tyrosine Kinase C-Terminal Src Kinase,Protein-Tyrosine Kinase c-src,Tyrosine Protein Kinase p50csk,Tyrosine-Protein Kinase CSK,c-src Kinase,c-src Tyrosine Kinase,CSK-src,p50(csk),C Terminal Src Kinase,CSK Tyrosine Protein Kinase,CSK src,CSK, Tyrosine-Protein Kinase,Carboxy Terminal Src Kinase,Kinase CSK, Tyrosine-Protein,Kinase c-src, Protein-Tyrosine,Kinase, C-Terminal Src,Kinase, CSK Tyrosine-Protein,Kinase, Carboxy-Terminal Src,Kinase, c-src,Kinase, c-src Tyrosine,Protein Tyrosine Kinase C Terminal Src Kinase,Protein Tyrosine Kinase c src,Src Kinase, C-Terminal,Src Kinase, Carboxy-Terminal,Tyrosine Kinase, c-src,Tyrosine Protein Kinase CSK,Tyrosine-Protein Kinase, CSK,c src Kinase,c src Tyrosine Kinase,c-src, Protein-Tyrosine Kinase
D018336 Receptors, Aryl Hydrocarbon Cytoplasmic proteins that bind certain aryl hydrocarbons, translocate to the nucleus, and activate transcription of particular DNA segments. AH receptors are identified by their high-affinity binding to several carcinogenic or teratogenic environmental chemicals including polycyclic aromatic hydrocarbons found in cigarette smoke and smog, heterocyclic amines found in cooked foods, and halogenated hydrocarbons including dioxins and polychlorinated biphenyls. No endogenous ligand has been identified, but an unknown natural messenger with a role in cell differentiation and development is suspected. AH Receptors,Aryl Hydrocarbon Receptors,Dioxin Receptors,Receptors, AH,Receptors, Dioxin,TCDD Receptors,AH Receptor,Aryl Hydrocarbon Receptor,Dioxin Receptor,Polyaromatic Hydrocarbon Receptor,Polyaromatic Hydrocarbon Receptors,Receptors, 2,3,7,8-Tetrachlorodibenzo-p-dioxin,Receptors, Polyaromatic Hydrocarbon,Receptors, TCDD,TCDD Receptor,Receptor, AH,Receptor, Aryl Hydrocarbon,Receptor, Dioxin,Receptor, Polyaromatic Hydrocarbon,Receptor, TCDD
D019061 src-Family Kinases A PROTEIN-TYROSINE KINASE family that was originally identified by homology to the Rous sarcoma virus ONCOGENE PROTEIN PP60(V-SRC). They interact with a variety of cell-surface receptors and participate in intracellular signal transduction pathways. Oncogenic forms of src-family kinases can occur through altered regulation or expression of the endogenous protein and by virally encoded src (v-src) genes. Protein-Tyrosine Kinases, src,Src Family Tyrosine Kinase,src Kinase,src Kinases,src Tyrosine Kinase,src-Family Kinase,src-Family Tyrosine Kinase,src Tyrosine Kinases,src-Family Tyrosine Kinases,Kinase, src,Kinase, src Tyrosine,Kinase, src-Family,Kinase, src-Family Tyrosine,Kinases, src,Kinases, src Protein-Tyrosine,Kinases, src Tyrosine,Tyrosine Kinase, src,Tyrosine Kinase, src-Family,Tyrosine Kinases, src,Tyrosine Kinases, src-Family,src Family Kinase,src Family Kinases,src Family Tyrosine Kinases,src Protein-Tyrosine Kinases

Related Publications

Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
July 2022, Journal of cellular biochemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
September 2003, The Journal of biological chemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
April 1998, Molecular pharmacology,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
December 2015, Toxicological research,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
August 1996, The Journal of biological chemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
April 1999, The Journal of biological chemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
November 2022, The Journal of biological chemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
November 2007, Molecular pain,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
September 2003, The Journal of biological chemistry,
Bin Dong, and Wei Cheng, and Wen Li, and Jie Zheng, and Dalei Wu, and Fumio Matsumura, and Christoph Franz Adam Vogel
July 2017, Structure (London, England : 1993),
Copied contents to your clipboard!