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Integrated global proteomic and phosphoproteomic analysis of cisplatin-induced apoptosis in A549 cells
Luyu Qi1, Jun Zhu2, Zhongyi Cheng2
1College of Life Sciences, Key Laboratory for Cell and Gene Engineering of Zhejiang Province, Zhejiang University, Hangzhou, China; Jingjie PTM Biolab (Hangzhou) Co. Inc, Hangzhou, China.
Abstract:
Protein phosphorylation, a widely occurring and significant post-translational modification, is integral to various biological processes. We previously utilized a protein affinity probe to identify genes damaged by cisplatin, revealing that it inflicts substantial damage on protein kinase and protein phosphatase genes. In this study, we investigated cisplatin-induced alterations in the global proteome and phosphoproteome of A549 cells. Employing Fe-IMAC beads and tyrosine phosphorylation enrichment antibodies, we identified 6944 protein groups and 18,274 phosphorylation sites on 4915 proteins across three biological replicates of both cisplatin-treated A549 cells and control cells. Among these, 730 tyrosine phosphorylation sites were identified-marking the most substantial discovery of such sites in A549 cells following cisplatin treatment. Bioinformatics analysis indicated that the proteins exhibiting significant phosphorylation level changes predominantly involved in RNA processing, modification, transcription, translation, and the spliceosome. This suggests that cisplatin-induced damage to protein kinases and phosphatases may disrupt the normal function of these proteins, consequently impairing DNA replication, RNA translation, and shearing, ultimately culminating in tumor cell death. Moreover, we cross-referenced our proteomic data with our previously obtained cisplatin-damaged genes, observing that the majority of down-regulated proteins derived from cisplatin-induced gene damage. The data are available on ProteomeXchange under the identifier PXD053902.
Insights
Cisplatin damages protein kinases and phosphatases, altering global cell phosphorylation. This disrupts RNA processing and translation, leading to tumor cell death and providing new insights into cisplatin
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Protein phosphorylation is a critical post-translational modification regulating biological processes.
- Cisplatin, a chemotherapy drug, is known to damage protein kinase and phosphatase genes.
Purpose of the Study:
- To investigate cisplatin's impact on the global proteome and phosphoproteome in A549 lung cancer cells.
- To identify specific phosphorylation changes induced by cisplatin treatment.
Main Methods:
- Utilized Fe-IMAC beads and tyrosine phosphorylation enrichment antibodies for proteomic and phosphoproteomic analysis.
- Analyzed global protein and phosphorylation site changes in cisplatin-treated versus control A549 cells.
- Performed bioinformatics analysis to identify affected biological pathways.
Main Results:
- Identified 6944 protein groups and 18,274 phosphorylation sites on 4915 proteins.
- Discovered 730 novel tyrosine phosphorylation sites in cisplatin-treated A549 cells.
- Found significant alterations in proteins involved in RNA processing, transcription, translation, and the spliceosome.
Conclusions:
- Cisplatin-induced damage to kinases/phosphatases disrupts essential cellular processes like RNA metabolism and translation.
- These disruptions contribute to impaired DNA replication and RNA function, ultimately causing tumor cell death.
- The study links cisplatin's proteomic effects to previously identified gene damage, highlighting its multifaceted impact on cancer cells.
