Dergi
/
Biyokimya
/
A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
JoVE Journal
Biyokimya
Bu içeriği görüntülemek için JoVE aboneliği gereklidir.  Oturum açın veya ücretsiz deneme sürümünü başlatın.
JoVE Journal Biyokimya
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

•

09:40 min

•

April 28, 2022

•

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

Bölümler

  • 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

Özet

Otomatik Çeviri

Otomatik Çeviri

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.

Etiketler

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

Article

Embed

ÇALMA LİSTESİNE EKLE

Usage Statistics

İlgili Videolar

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

A Tandem Liquid Chromatography–Mass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions

Protein Digestion, Ultrafiltration, and Size Exclusion Chromatography to Optimize the Isolation of Exosomes from Human Blood Plasma and Serum

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Untargeted Liquid Chromatography-Mass Spectrometry-Based Metabolomics Analysis of Wheat Grain

Read Article