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PJ综合征(Peutz-Jeghers syndrome,PJS)的疾病基因STK11编码由433个氨基酸残基组成的丝氨酸/苏氨酸蛋白激酶STK11,具有调控细胞周期,调节细胞极性以及细胞基础能量代谢等重要功能.STK11蛋白羧基端含有123个残基,对STK11生物学功能的实现起着调节作用,但机制尚未明确.本研究利用pGEX4T-2原核表达系统,构建pGEX4T-2-STK11羧基端重组载体,并诱导表达了GST-STK11羧基端融合蛋白,采用GST-pulldown结合质谱分析的方法,鉴定出STK11羧基端的可能互作蛋白LOH12CR1.通过免疫共沉淀技术证实了STK11激酶同LOH12CR1互作;免疫荧光共定位实验亦发现STK11与LOH12CR1共定位.STK11与LOH12CR1互作的发现为研究STK11的功能提供新的切入点.
STK11, a disease gene of Peutz-Jeghers syndrome (PJS), encodes a serine / threonine protein kinase STK11 consisting of 433 amino acid residues and has important functions of regulating cell cycle, regulating cell polarity and basal energy metabolism of cells The carboxyl terminal of STK11 protein contains 123 residues, which plays a regulatory role in the biological function of STK11, but the mechanism is not yet clear.In this study, the pGEX4T-2 prokaryotic expression system was used to construct pGEX4T-2-STK11 carboxyl terminal recombinant vector The GST-STK11 carboxyl terminal fusion protein was induced and expressed, and the possible interaction protein LOH12CR1 at the carboxyl terminal of STK11 was identified by GST-pulldown combined with mass spectrometry analysis.The co-localization of STK11 kinase and LOH12CR1 was confirmed by co-immunoprecipitation technique.Immunofluorescence co-localization The experiment also found that co-localization of STK11 and LOH12CR1.The discovery of the interaction between STK11 and LOH12CR1 provided a new entry point for studying the function of STK11.