State Key Laboratory of Antiviral Drugs, Pingyuan Laboratory, NMPA Key Laboratory for Research and Evaluation of Innovative Drug, School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, Henan, China.
Department of Nephrology and Rheumatology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Methods Mol Biol. 2025;2854:29-34. doi: 10.1007/978-1-0716-4108-8_4.
Mass spectrometers are widely used to identify protein phosphorylation sites. The process usually involves selective isolation of phosphoproteins and subsequent fragmentation to identify both the peptide sequence and phosphorylation site. Immunoprecipitation could capture and purify the protein of interest, greatly reducing sample complexity before submitting it for mass spectrometry analysis. This chapter describes a method to identify an abnormal phosphorylated site of the adaptor protein by a viral kinase through immunoprecipitation followed by LC-MS/MS.
质谱仪广泛用于鉴定蛋白质磷酸化位点。该过程通常涉及磷酸化蛋白的选择性分离,随后进行片段化以鉴定肽序列和磷酸化位点。免疫沉淀可以捕获和纯化感兴趣的蛋白质,在进行质谱分析之前大大降低了样品的复杂性。本章描述了一种通过免疫沉淀结合 LC-MS/MS 方法鉴定病毒激酶引起的衔接蛋白异常磷酸化位点的方法。