Proteomic Analysis of Downstream Proteins Regulated by THRAP3 in THP-1 Cells

This study aimed to analyze the downstream proteins regulated by THRAP3 in THP-1 cells using proteomics. THP-1 cells were divided into control and KO groups (with 3 biological replicates each, transfected with sgNC or sgTHRAP3 respectively). After cell lysis, trypsin digestion, Tandem Mass Tags labeling, fractionation, and resuspension, peptide fractions were separated via LC and analyzed by MS on a Q-Exactive HF Orbitrap mass spectrometer. MS data were interpreted with Percolator v.355 to identify UniProt sequences with a 1% FDR.

本研究旨在通过蛋白质组学技术分析THP-1细胞中受THRAP3调控的下游蛋白。将THP-1细胞分为对照组与敲除（KO）组，每组各设置3次生物学重复，分别转染sgNC或sgTHRAP3。经细胞裂解、胰蛋白酶酶解、串联质量标签（Tandem Mass Tags）标记、组分分级分离与重悬后，通过液相色谱（Liquid Chromatography, LC）分离肽段组分，并在Q-Exactive HF Orbitrap质谱仪上进行质谱（Mass Spectrometry, MS）分析。使用Percolator v.355软件解析质谱数据，以1%的错误发现率（False Discovery Rate, FDR）鉴定UniProt数据库序列。