Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Proton Transfer Charge Reduction Enables Isobaric Labeling-Based Proteoform Quantification of Overlapping Signals in Top-Down Mass Spectrometry.

Journal of the American Society for Mass Spectrometry·2026
Same author

ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.

Journal of extracellular biology·2026
Same author

H<sub>2</sub>-dependent modulation of tetrahydromethanopterin S-methyltransferase (Mtr complex) activity by the small protein MtrR in Methanosarcina mazei.

The FEBS journal·2026
Same author

CoMPaseD: advanced planning of proteomic experiments aiming to identify small proteins.

microLife·2026
Same author

An ancient lysozyme in placozoans participates in acidic extracellular digestion.

Communications biology·2026
Same author

Meprin β elevates hippocampal soluble Aβ in the APP/V717I mouse model.

Experimental neurology·2025
Same journal

From Method-Defined Signals to Reference Measurement Procedures: Two Decades of Mass Spectrometry-Based ProGRP Quantification.

Journal of proteome research·2026
Same journal

Proteomic Profiling of Extracellular Vesicle-Enriched Plasma Using Mag-Net for Biomarker Discovery in Pancreatic Ductal Adenocarcinoma.

Journal of proteome research·2026
Same journal

Computationally Efficient Bayesian Estimation of Graphical Networks for Omics Data.

Journal of proteome research·2026
Same journal

Hierarchy of MS-Based Evidence.

Journal of proteome research·2026
Same journal

Proteomic Profiling of Exosomes from HPV-Positive and HPV-Negative Head and Neck Squamous Cell Carcinoma: Selective Cargo Packaging.

Journal of proteome research·2026
Same journal

Proteomic Analysis Identifies ATE1-Dependent Arginylation Dysregulation across Meningioma Grades.

Journal of proteome research·2026
See all related articles

Related Experiment Video

Updated: May 11, 2026

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
12:23

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

Published on: August 2, 2018

Optimized fragmentation conditions for iTRAQ-labeled phosphopeptides.

Dennis Linke1, Chien-Wen Hung, Liam Cassidy

  • 1AG Systematische Proteomforschung & Bioanalytik - Institut für Experimentelle Medizin, Christian-Albrechts-Universität zu Kiel, 24105 Kiel, Germany.

Journal of Proteome Research
|May 15, 2013
PubMed
Summary
This summary is machine-generated.

This study optimizes mass spectrometry methods for quantifying phosphorylated proteins using isobaric labeling. An improved protocol balances identification and quantification, enabling efficient analysis of complex biological samples.

More Related Videos

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
12:11

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Published on: February 27, 2020

Covalent Fragment Screening Using the Quantitative Irreversible Tethering Assay
06:17

Covalent Fragment Screening Using the Quantitative Irreversible Tethering Assay

Published on: February 28, 2025

Related Experiment Videos

Last Updated: May 11, 2026

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
12:23

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

Published on: August 2, 2018

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
12:11

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Published on: February 27, 2020

Covalent Fragment Screening Using the Quantitative Irreversible Tethering Assay
06:17

Covalent Fragment Screening Using the Quantitative Irreversible Tethering Assay

Published on: February 28, 2025

Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Post-Translational Modifications

Background:

  • Protein phosphorylation is a key regulatory mechanism in biological processes.
  • Simultaneous identification, localization, and quantification of phosphoproteins are crucial for understanding cellular signaling.
  • Isobaric labeling strategies like iTRAQ are used for quantitative phosphopeptide analysis.

Purpose of the Study:

  • To evaluate different MS/MS fragmentation modes (CID, CID + MSA, HCD) on the Orbitrap Velos MS for quantitative phosphopeptide analysis.
  • To determine an optimized MS/MS protocol for simultaneous identification and quantification of iTRAQ-labeled phosphopeptides.
  • To assess the performance of fragmentation modes in semicomplex and complex phosphopeptide mixtures.

Main Methods:

  • Utilized Orbitrap Velos MS with Collision Induced Dissociation (CID), CID plus Multistage Activation (MSA), and Higher Energy Collision Dissociation (HCD) fragmentation.
  • Analyzed semicomplex (12) and complex (131) phosphopeptide mixtures labeled with iTRAQ.
  • Optimized normalized collision energies for balancing peptide identification and quantification.

Main Results:

  • A compromise in collision energies is needed for simultaneous quantification and identification of iTRAQ-labeled phosphopeptides.
  • An optimized protocol involving sequential CID (for identification) and HCD (for parallel identification/quantification) was developed.
  • This protocol meets the time constraints of Liquid Chromatography-Mass Spectrometry (LC-MS/MS) experiments.

Conclusions:

  • The developed MS/MS protocol enhances the efficiency of quantitative phosphoproteomics.
  • Optimized fragmentation strategies enable robust identification and quantification of phosphopeptides.
  • This method facilitates a deeper understanding of dynamic regulatory roles of protein phosphorylation in biological systems.