摘要
目的鉴定端粒重复结合蛋白1(TRF1)的磷酸化位点及其与磷酸化激酶Aurora B在体内外的相互作用。方法用myc-His-TRF1真核表达质粒转染人宫颈癌细胞HeLa,免疫沉淀后行质谱分析。采用免疫沉淀及体外沉降实验研究TRF1与Aurora B的相互作用。结果免疫沉淀实验结合质谱分析鉴定出5个新的TRF1磷酸化位点;运用体外沉降实验和体内免疫共沉淀实验证实Aurora B能够与TRF1在体内外相互结合。结论Aurora B可能是一个新的TRF1结合蛋白,TRF1与Aurora B能够在体内外相互结合。
Objective To determine the phosphorylation sites of telomeric repeat binding factor 1(TRF1)and analyze the interaction of TRF1 and phosphorylation kinase Aurora B in vitro and in vivo.Methods The eukaryotic expression plasmid myc-His-TRF1 was transfected into human HeLa cells,and then was identified by mass spectrometry after immunoprecipitation.The interaction of TRF1 and Aurora B was studied by immunoprecipitation assay and protein pull-down assay in vitro.Results Five new phosphorylation sites of TRF1 were discovered with immunoprecipitation and mass spectrometry assay.Protein pull-down assay in vitro and co-immunoprecipitation assay in vivo confirmed that Aurora B and TRF1 could bind to each other in vivo and in vitro.Conclusion Aurora B may be a new TRF1 binding protein.TRF1 and Aurora B can bind to each other in vivo and in vitro.
作者
王冲
李玲玲
李梦亚
申晓辉
王树娟
WANG Chong;LI Lingling;LI Mengya;SHEN Xiaohui;WANG Shujuan(Department of Hematology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China)
出处
《新乡医学院学报》
CAS
2022年第1期6-8,共3页
Journal of Xinxiang Medical University
基金
国家自然科学基金资助项目(编号:U1804191、U1904137)
河南省重点研发与推广专项(科技攻关)(编号:182102310563)。