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Data-Independent Acquisition (DIA)-Based Label-Free Redox Proteomics (DIALRP) Identifies Prominent Cysteine Oxidations in Translation Machinery in Prostate Cancer Cells Under Oxidative Stress

  • 0Department of Omics and Systems Biology, Graduate School of Medical and Dental Sciences, Niigata University, 757 Ichibancho, Asahimachi-dori, Chuo-ku, Niigata City, Niigata 951-8510, Japan.
Journal of Proteome Research +

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July 31, 2025

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July 31, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

Oxidative stress alters proteins, impacting cell function. This study introduces a new method to track these changes, revealing how stress disrupts translation complexes in prostate cancer cells.

Area Of Science

  • Proteomics
  • Cellular Biology
  • Biochemistry

Background

  • Oxidative stress significantly impacts physiological and pathological processes, including aging, cancer, and neurodegenerative diseases.
  • Protein cysteine residues are highly vulnerable to oxidative modifications, altering protein function and intracellular signaling.
  • Understanding these modifications is crucial for deciphering cellular responses to stress.

Purpose Of The Study

  • To develop and apply a novel data-independent acquisition mass spectrometry (DIA-MS)-based redox proteomics method (DIALRP) for comprehensive analysis of cysteine oxidation.
  • To investigate the impact of menadione (MND)-induced oxidative stress on protein cysteine modifications in the DU145 prostate cancer cell line.
  • To elucidate the functional consequences of oxidative stress on cellular processes, particularly translation regulation.

Main Methods

  • Utilized a label-free, data-independent acquisition mass spectrometry (DIA-MS) approach named DIALRP.
  • Applied DIALRP to analyze cysteine oxidative modifications in DU145 cells treated with menadione (MND).
  • Performed quantitative analysis of redox changes in cysteine-containing peptides and gene ontology analysis.

Main Results

  • Identified 10,821 cysteine-containing peptides and quantified redox changes in 3665 peptides.
  • Observed significant oxidation in 1407 peptides upon MND treatment.
  • Found enrichment of translation-related molecules among sensitive proteins and demonstrated inhibition of EIF2, EIF6, and EEF2 complex formation.

Conclusions

  • DIALRP is a robust and cost-effective method for studying redox-regulated cellular processes.
  • MND-induced oxidative stress inhibits key translation initiation factors, leading to reduced translation activity.
  • These findings offer critical insights into translation regulation under oxidative stress and provide a foundation for future redox biology research.

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