JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE) ›› 2021, Vol. 41 ›› Issue (1): 1-7.doi: 10.3969/j.issn.1674-8115.2021.01.001

• Basic research • Previous Articles     Next Articles

Phosphoproteomic analysis of concanavalin A-induced hepatocytes injury

Zhuo-yi YANG1(), Hui CHEN1, Si-yi BAI1, Pameila PERHATI1, Jing-li HOU2, Yun-sheng YUAN1()   

  1. 1.Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education; Shanghai Jiao Tong University School of Pharmacy, Shanghai 200240, China
    2.Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai 200240, China
  • Online:2021-01-28 Published:2021-02-22
  • Contact: Yun-sheng YUAN E-mail:yangzhuoyi@sjtu.edu.cn;yunsheng@sjtu.edu.cn
  • Supported by:
    Funding Information] National Science and Technology Major Project(2019ZX09201001);National Natural Science Foundation of China(31671388);Shanghai Jiao Tong University Medical-Engineering Joint Project(YG2019QNA50)

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Abstract: Objective

·To study the changes of phosphorylation level of whole-cell proteins during the process of concanavalin A (ConA)-induced hepatocyte injury.

Methods

·Mouse hepatocytes of AML12 cells and ConA (10 μg/mL) were co-cultured to induce a hepatocyte injury model. After 12 h, the total proteins were extracted and digested into peptides. The peptides were enriched by metal oxide affinity chromatography and detected by using high performance liquid chromatography-mass spectrometer. Identification and relative quantitative analysis of phosphorylation sites and peptides were performed using the Label free method. Proteins with a difference of more than two times of phosphorylation level of each site were screened. And the biological function enrichment analysis and protein interaction regulation network analysis were performed.

Results

·In the ConA-induced AML12 cell injury model, a total of 11 200 phosphorylated modified peptides and 2 685 localizable and quantifiable peptides were identified. Among them, there were 82 differentially regulated phosphorylated modified peptides, corresponding to 77 proteins. These phosphorylated proteins were mainly related to protein binding and enzyme activities. And the main pathways were related to apoptosis and RAS kinase pathways.

Conclusion

·AML12 cells undergo phosphorylation level change in ConA-induced liver cell injury and these differentially phosphorylated proteins may be related to liver cell apoptosis.

Key words: concanavalin A (ConA), AML12 cell line, proteomics, phosphorylation modification

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