Identification of membrane proteins in Chinese hamster ovary cell using Orbitrap fusion lumos mass spectrometers based on non-label technique

doi:10.19871/j.cnki.xfcrbzz.2024.02.007

Abstract

Abstract: Objective To explore the method of identifying the whole proteome and membrane protein of Chinese hamster ovary (CHO) cells by using the Orbitrap fusion lumos mass spectrometer based on label-free technique,and evaluate its application effect. It provides the methodological basis for studying the pathogenesis of membrane protein-related infectious diseases. Method CHO cells were cultured, and then the whole protein and membrane protein of the cells in the logarithmic growth phase were extracted, enriched and enzymatically hydrolyzed. The peptides were identified by HPLC-Orbitrap fusion lumos three in mass spectrometer. Maxquant software and Uniprot database were used for database search and identification. Finally, the identified membrane proteins were classified by GO function. Result A total of 14 300 peptide segments were detected in the whole proteome, and 2708 proteins were found in the database search,including 2002 credible proteins(FDR<1% and unique peptide>1); A total of 13 179 peptide segments were detected in the membrane proteome, and 2594 proteins were found in the database search,including 1873 credible proteins. Among the top 20 proteins in the identified total protein and membrane protein groups, there are 4 membrane related proteins in the total protein group, accounting for 20%; There are 10 membrane related proteins in the membrane proteome, accounting for 50%. In the membrane protein components. GO function classification shows that molecular function is mainly related to binding and catalytic activity, while in biological processes, it mainly involves cellular processes, metabolic processes. Conclusion The membrane-associated proteins in the membrane protein group were significantly higher than those in the whole protein group,and the application of Orbitrap fusion lumos analysis technology based on label-free technique is an effective method for identifying cell membrane proteins.

Key words: Chinese hamster ovary cells, Membrane protein, Non labeling method, Orbitrap fusion high-resolution mass spectrometer

CLC Number:

Liu Yawen, Liang Xiongshun, Hu Xuqiao, Liu Xiaoqian, Xu Wanna, Hong Wenxu. Identification of membrane proteins in Chinese hamster ovary cell using Orbitrap fusion lumos mass spectrometers based on non-label technique[J]. Electronic Journal of Emerging Infectious Diseases, 2024, 9(2): 34-40.

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