双分子荧光互补
互补
蛋白质片段互补分析
激酶
蛋白质-蛋白质相互作用
细胞生物学
蛋白激酶A
生物化学
生物
化学
生物物理学
基因
突变体
作者
Mayu Kamimura,Yulong Han,Nobuki Kito,Fang‐Sik Che
标识
DOI:10.1080/09168451.2014.882757
摘要
Abstract Protein kinases are key regulators of cell function that constitute one of the largest and most functionally diverse gene families. We developed a novel assay system, based on the bimolecular fluorescence complementation (BiFC) technique in Escherichia coli, for detecting transient interactions such as those between kinases and their substrates. This system detected the interaction between OsMEK1 and its direct target OsMAP1. By contrast, BiFC fluorescence was not observed when OsMAP2 or OsMAP3, which are not substrates of OsMEK1, were used as prey proteins. We also screened for interacting proteins of calcium-dependent protein kinase 8 (OsCPK8), a regulator of plant immune responses, and identified three proteins as interacting molecules of OsCPK8. The interaction between OsCPK8 and two of these proteins (ARF-GEF and peptidyl prolyl isomerase) was confirmed in rice cells by means of BiFC technology. These results indicate that our new assay system has the potential to screen for protein kinase target molecules.
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