Journal
/
Biochemistry
/
LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

DOI:
10.3791/55626-v

•

08:01 min

•

April 09, 2017

•

, ,
1Division of Structural Biology and Biochemistry, School of Biological Sciences,Nanyang Technological University

Chapters

  • 00:05Title
  • 00:44Long-length Anion-exchange (LAX) Capillary Column Packing
  • 01:43Tissue Sample Preparation
  • 05:33One-dimension Long-length Electrostatic Repulsion-hydrophilic Interaction Chromatography-tandem Mass Spectrometry (LERLIC-MS/MS) Separation and Data Analysis
  • 06:48Results: Separation of Deamidation Isoforms of Glutamine and Asparagine by LERLIC-MS/MS
  • 07:34Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present a step-by-step protocol of the long-length electrostatic repulsion-hydrophilic interaction chromatography-tandem mass spectrometry (LERLIC-MS/MS) method. This is a novel methodology that enables for the first time quantification and characterization of the glutamine and asparagine deamidation isoforms by shotgun proteomics.

Tags

Keywords: LERLIC-MS/MSGlutamine DeamidationAsparagine DeamidationShotgun ProteomicsPeptide SeparationAnion Exchange ChromatographyProtein QuantificationDithiothreitolIodoacetamide

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/55626?article_type=v

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
check_url/55626?article_type=v

Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
check_url/55626?article_type=v

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
check_url/55626?article_type=v

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
check_url/55626?article_type=v

SUMO-Binding Entities (SUBEs) as Tools for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Cancer
check_url/55626?article_type=v

TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis
check_url/55626?article_type=v

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
check_url/55626?article_type=v

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics
check_url/55626?article_type=v

Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot
check_url/55626?article_type=v

Shotgun Lipidomics of Rodent Tissues

Read Article