Malonylation Proteomics Study of Hepatocellular Carcinoma Tissues Using Liquid Chromatography-Mass Spectrometry
10.19756/j.issn.0253-3820.251158
- VernacularTitle:基于液相色谱-质谱联用技术的肝癌组织的丙二酰化修饰组学研究
- Author:
Meng ZHANG
1
;
Xiao LONG
;
Yan-Nan WU
;
Hao SI
;
Hong-Xia WANG
Author Information
1. 宁波大学材料科学与化学工程学院,宁波 315211;宁波镇海质谱研究院,宁波 315211
- Keywords:
Hepatocellular carcinoma;
Malonylation proteomics;
Liquid chromatography-mass spectrometry;
Antibody-based affinity enrichment
- From:
Chinese Journal of Analytical Chemistry
2025;53(9):1546-1554
- CountryChina
- Language:Chinese
-
Abstract:
Malonylation is an important post-translational modification of proteins.In this work,a comprehensive malonylation proteomics study on hepatocellular carcinoma(HCC)tumorous and non-tumorous tissues using antibody enrichment combined with high performance liquid chromatography-mass spectrometry for discovery of early diagnostic biomarkers or potential new drug targets of HCC was performed.A total of 1299 malonylated peptides containing 1064 malonylated sites were identified from HCC tissues,corresponding to 511 malonylated proteins.Quantitative results showed that 56 and 80 malonylated proteins were up-regulated and down-regulated in HCC tissues,including 60 and 101 malonylated sites,respectively.Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis showed that these differentially modified proteins were involved in various important pathways such as metabolic pathways,fatty acid degradation,and glycolysis/gluconeogenesis.As a key enzyme in glycolysis/gluconeogenesis,phosphoenolpyruvate carboxykinase 1(PCK1)was malonylated at lysine 244(K244)and the malonylation was only detected in HCC tumorous tissues.More importantly,the K244 site served as a binding site for Mn2+and highly conserved across different species.Therefore,it could speculate that the malonylation of K244 would affect its activity and played a role in liver cancer by affecting its binding with Mn2+,which requied further verification through site mutation experiments.Western blot analysis by malonylation pan antibody showed that the malonylation level reduced markedly in HCC tumorous tissues compared with adjacent non-tumorous tissues,which was consistent with mass spectrometry data.In addition,the proliferation and invasion of PLC/PRF/5 cell was significantly inhibited and protein malonylation level was increased obviously when treated with sodium malonate.All the evidence indicated that protein malonylation played an important role in HCC pathogenesis,and its molecular mechanism deserved further investigation.Furthermore,the 136 differentially malonylated proteins provided rich source of candidate targets for further research on HCC pathogenesis.
