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A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
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Biochemistry
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JoVE Journal Biochemistry
A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

DOI:
10.3791/62409-v

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09:40 min

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April 28, 2022

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, , ,
1Department of Experimental Oncology,European Institute of Oncology IRCCS (IEO), 2European School of Molecular Medicine (SEMM), c/o Campus IFOM-IEO

Chapters

  • 00:04Introduction
  • 00:44Lysate Digestion
  • 02:08Off‐Line HpH‐RP Chromatographic Fractionation of Peptides
  • 02:53R‐Methylated Peptide Immuno‐Affinity Enrichment
  • 06:47Results: R‐Methylation Analysis by Mass Spectrometry
  • 08:39Conclusion

Summary

Automatic Translation

Automatic Translation

Protein Arginine (R)-methylation is a wide-spread post-translational modification regulating multiple biological pathways. Mass spectrometry is the best technology to globally profile the R-methyl-proteome, when coupled to biochemical approaches for modified peptide enrichment. The workflow designed for the high confidence identification of global R-methylation in human cells is described here.

Tags

Keywords: Mass SpectrometryProteomicsProtein R-methylationPRMT InhibitorsDTTIodoacetamideTrypsinLysargiNaseHigh PH Reversed Phase Liquid ChromatographyImmuno-affinity Enrichment

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