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用于活细胞中ERK2二聚体的基因编码生物发光指示剂。

Genetically encoded bioluminescent indicator for ERK2 dimer in living cells.

作者信息

Kaihara Asami, Umezawa Yoshio

机构信息

Department of Chemistry, School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

出版信息

Chem Asian J. 2008 Jan 4;3(1):38-45. doi: 10.1002/asia.200700186.

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 17beta-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.

摘要

在本研究中,采用分裂型海肾荧光素酶互补法开发了一种用于ERK2二聚体的基因编码生物发光指示剂,其中ERK2二聚体的形成在活细胞中诱导生物发光的自发发射。响应于细胞外刺激,如表皮生长因子(EGF)或17β-雌二醇(E2),细胞外信号调节激酶2(ERK2)被其上游激酶MEK磷酸化,并且还在细胞的各个区域(包括细胞核)中使其底物磷酸化。磷酸化的ERK2导致形成其二聚体,从而将自身转运到细胞核中。我们用该指示剂证明,EGF或E2刺激可诱导活的MCF-7细胞中ERK2二聚体的形成。对这种二聚体形成的动力学进行了研究和讨论。

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