Genetically Encoded Bioluminescent Indicator for ERK2 Dimer in Living Cells |
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| Authors: | Asami Kaihara Dr. Yoshio Umezawa Prof. Dr. |
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| Affiliation: | 1. Department of Chemistry, School of Science, The University of Tokyo, Hongo, Bunkyo‐ku, Tokyo 113‐0033, Japan, Fax: (+81)?42 468 9292;2. Current address: Medical Research Institute, Tokyo Medical and Dental University, Kandasurugadai, Chiyoda‐ku, Tokyo 101‐0062, Japan;3. Current address: Research Institute of Pharmaceutical Sciences, Musashino University, Shinmachi, Nishitokyo‐shi, Tokyo, 202‐8585, Japan |
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| Abstract: | In this study, a genetically encoded bioluminescent indicator for ERK2 dimer was developed with the split Renilla luciferase complementation method, in which the formation of ERK2 dimer induces a spontaneous emission of bioluminescence in living cells. In response to extracellular stimuli, such as epidermal growth factor (EGF) or 17β‐estradiol (E2), extracellular signal‐regulated kinase 2 (ERK2) is phosphorylated by its upstream kinase MEK, and also phosphorylates its substrates in various regions of the cell, including the nucleus. Phosphorylated ERK2 is led to form its dimer, thereby transporting itself into the nucleus. We demonstrated with the indicator that stimulation with EGF or E2 induces the formation of ERK2 dimer in living MCF‐7 cells. The dynamics of this dimer formation was examined and discussed. |
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| Keywords: | biosensors dimerization ERK luminescence phosphorylation |
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