Comprehensive profiling of CTP-binding proteins using a biotinylated CTP affinity probe  

作　　者：Mengting Pa Yunming Liu Xiaofang Zheng Meijuan Zhou Changjun You Xiaoxia Dai 

机构地区：[1]State Key Laboratory of Chemo/Biosensing and Chemometrics,College of Chemistry and Chemical Engineering,Molecular Science and Biomedicine Laboratory,Hunan Provincial Key Laboratory of Biomacromolecular Chemical Biology,Hunan University,Changsha 410082,China

出　　处：《Chinese Chemical Letters》2021年第11期3479-3482,共4页中国化学快报（英文版）

基　　金：supported by the National Natural Science Foundation of China(Nos.21807030,21907028);the Science and Technology Innovation Program of Hunan Province(No.2019RS2020);Natural Science Foundation of Hunan Province(No.2020JJ5046);the Fundamental Research Funds for the Central Universities(Nos.531118010061,531118010259).

摘　　要：Recent studies have shown that CTP may act as a ligand to regulate the activity of its target proteins in many biological processes.However,proteome-wide identification of CTP-binding proteins remains challenging.Here,we employed a biotinylated CTP affinity probe coupled with stable isotope labeling by amino acids in cell culture(SILAC)-based quantitative proteomics approach to capture,identify and quantify CTP-binding proteins in human cells.By performing two types of competitive SILAC experiments with high vs.low concentrations of CTP probe(100 vs.10µmol/L)or with CTP probe in the presence of free CTP,we identified 90 potential CTP-binding proteins which are involved in a variety of biological processes,including protein folding,nucleotide binding and cell-cell adhesion.Together,we developed a chemical proteomic method for uncovering the CTP-binding proteins in human cells,which could be widely applicable for profiling CTP-binding proteins in other biological samples.

关 键 词：CTP-binding proteins CTP affinity probe Mass spectrometry SILAC Quantitative proteomics 

分 类 号：R313[医药卫生—基础医学]