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Global Phosphoproteomic Analysis Reveals Significant Metabolic Reprogramming in the Termination of Liver Regeneration in Mice
journal contribution
posted on 2020-03-10, 18:22 authored by Jingzi Zhang, Neng Tang, Yinjuan Zhao, Ruoyu Zhao, Xiao Fu, Dandan Zhao, Yue Zhao, Lan Huang, Chaojun Li, Yudong Qiu, Bin Xue, Lei FangPhosphorylation is crucial in regulating various biological processes.
However, comprehensive phosphoproteomic profiling in the termination
of liver regeneration (LR) is still missing. Here, we used Tandem
Mass Tag (TMT) labeling coupled with phosphopeptide enrichment and
two-dimensional (2D) liquid chromatography–mass spectrometry
(LC–MS)/MS analysis to establish a global phosphoproteomic
map in the liver of mice at day 5 after partial hepatectomy (PH).
Altogether, 9731 phosphosites from 3443 proteins were identified and
7802 phosphosites from 2980 proteins were quantified. Motif analysis
of the identified phosphosites revealed a diverse array of consensus
sequences, suggesting that multiple kinase families including ERK/MAPK,
PKA/PKC, CaMK-II, CKII, and CDK may be involved in the termination
of LR. Functional clustering analysis of proteins with dysregulated
phosphosites showed that they mainly participate in metabolic pathways,
DNA replication, and tight junction. More importantly, the deletion
of PP2Acα in the liver remarkably changes the overall phosphorylation
profile, indicating its critical role in regulating the termination
of LR. Finally, several differentially phosphorylated sites were validated
by co-immunoprecipitation and Western blot. Taken together, our data
unravel the first comprehensive phosphoproteomic map in the termination
of LR in mice, which greatly expands our knowledge in the complicated
regulation of this process and provides new directions for the treatment
of liver cancer using liver resection.