Actinomycin D identified as an inhibitor of discoidin domain receptor 2 interaction with collagen through an insect cell based screening of a drug compound library.
Discoidin domain receptors belong to the cell surface receptor tyrosine kinase family and recognize collagens for their activating ligands. They have been implicated for cell growth and migration and their elevated expressions were observed in various human cancers. When we expressed human Discoidin domain receptor 2 (DDR2) in insect cells, the protein was targeted properly into the cell membrane and this could enforce the cells to adhere on culture plate coated with type I collagen. By taking advantage of this, we established a novel insect cell based screening protocol to identify chemicals which inhibit the interaction between DDR2 and collagen. We screened a drug-compound library to select an anti-cancer drug, actinomycin D, as the inhibitory compound. Actinomycin D prevented the activation of DDR2 by type I collagen in human embryonic kidney 293 cells with an IC(50) value of 9 microM, while it did not interfere with the activation of other receptor tyrosine kinases by their ligands. In conclusion we identified a new biological function of actinomycin D and the insect cell based method provides a useful protocol for screening inhibitors against the association of DDR2 with collagen.
Siddiqui
Kauser
K
Biomedical Research Center, Korea Institute of Science and Technology, Seoul.
Kim
Geun Woong
GW
Lee
Dong Hoon
DH
Shin
Hae Ryun
HR
Yang
Eun Gyeong
EG
Lee
Nam Taek
NT
Yang
Beom-Seok
BS
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002461:Cell Line, Transformed; D024042:Collagen Type I / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology*; D000070818:Discoidin Domain Receptors; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D007313:Insecta; D019151:Peptide Library; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D024042:Collagen Type I; D019151:Peptide Library; D011975:Receptors, Mitogen; D011994:Recombinant Proteins; D014415:Tunicamycin; D003609:Dactinomycin; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases