中药与临床2025，Vol.16Issue(3)：16-21,50,7.
基于纳升液相色谱-高分辨质谱技术的僵蚕蛋白质组分析
Proteomic analysis of Bombyx batryticatus based on nano liquid chromatography-high resolution mass spectrometry
摘要
Abstract
Objective:To optimize the method of extracting the protein of Bombyx batryticatus suitable for liquid-mass spectrometry analysis,and to perform proteomic analysis of the extracted total protein of Bombyx batryticatus to get a comprehensive understanding of the protein composition of the Bombyx batryticatus.Methods:Firstly,The total proteins of Bombyx mori were extracted by RIPA(Method ①),T-PER(Method ②),and Tris-HCl extraction(Method ③),respectively.Secondly,the efficiencies of acetone precipitation,ultrafiltration-assisted digest(FASP),and the combination of the two on proteolysis were compared.Then the influences of the key instrumental parameters and the liquid-phase separation gradient on the mass spectrometry data acquisition were comparatively analyzed.After that,the optimized method was adopted for the proteome analysis of Bombyx batryticatus samples.The optimized method was finally adopted for the proteomic analysis of Bombyx batryticatus samples.Results:With DDA data scanning mode,total proteins of Bombyx batryticatus were extracted by Tris-HCl extraction and separated by 90 min nanoliter reverse liquid chromatography after acetone-FASP digestion,and the highest number of Bombyx batryticatus proteins were identified.On this basis,bioinformatics analysis of the identified proteins of Bombyx batryticatus showed that the proteins of Bombyx batryticatus were mainly hydrophilic and distributed mainly in the cytoplasm and nucleus.The majority of proteins have functional activities such as binding,translational transport,and catalysis,with the largest percentage of proteins involved in protein translation and the formation of translation initiation complexes.Conclusion:This study demonstrats that the identified proteins of Bombyx batryticatus have a wide range of biological functions,which lays the foundation for the study of the biological activities of the proteins in Bombyx batryticatus.关键词
僵蚕/高分辨质谱/蛋白提取/生物信息学Key words
Bombyx batryticatus/high resolution mass spectrometry/protein extraction/bioinformatics分类
医药卫生引用本文复制引用
张梦姣,王巍,李雪敏,段婷引,邓放..基于纳升液相色谱-高分辨质谱技术的僵蚕蛋白质组分析[J].中药与临床,2025,16(3):16-21,50,7.基金项目
四川省科技厅重点研发项目(2022YFN0068) （2022YFN0068）
