Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

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

High-Throughput Large-Scale Targeted Proteomics Assays for Quantifying Pathway Proteins in <i>Pseudomonas putida</i> KT2440.

Frontiers in bioengineering and biotechnology·2021

Same author

Online supercritical fluid extraction mass spectrometry (SFE-LC-FTMS) for sensitive characterization of soil organic matter.

Faraday discussions·2019

Same author

Circadian Proteomic Analysis Uncovers Mechanisms of Post-Transcriptional Regulation in Metabolic Pathways.

Cell systems·2018

Same author

Formularity: Software for Automated Formula Assignment of Natural and Other Organic Matter from Ultrahigh-Resolution Mass Spectra.

Analytical chemistry·2017

Same author

LC-SRM-Based Targeted Quantification of Urinary Protein Biomarkers.

Methods in molecular biology (Clifton, N.J.)·2017

Same author

Quantification of mutant SPOP proteins in prostate cancer using mass spectrometry-based targeted proteomics.

Journal of translational medicine·2017

Search research articles

Related Experiment Video

Updated: Jun 23, 2026

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Integrated workflow for characterizing intact phosphoproteins from complex mixtures.

Si Wu¹, Feng Yang, Rui Zhao

¹Pacific Northwest National Laboratory, Richland, Washington 99352, USA.

Analytical Chemistry

|May 12, 2009

Summary

This study introduces a novel metal-free liquid chromatography-mass spectrometry platform for analyzing intact phosphoproteins. This technology accurately maps multisite phosphorylation, offering a comprehensive view of cellular signaling states.

More Related Videos

Phosphoproteomic Strategy for Profiling Osmotic Stress Signaling in Arabidopsis

Phosphoproteomic Strategy for Profiling Osmotic Stress Signaling in Arabidopsis

Published on: June 25, 2020

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

Related Experiment Videos

Last Updated: Jun 23, 2026

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Phosphoproteomic Strategy for Profiling Osmotic Stress Signaling in Arabidopsis

Phosphoproteomic Strategy for Profiling Osmotic Stress Signaling in Arabidopsis

Published on: June 25, 2020

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

Published on: July 26, 2019

Area of Science:

Proteomics
Cellular Signaling
Biochemistry

Background:

Multisite phosphorylation is crucial for cellular signal integration and understanding protein function.
Current methods struggle to accurately quantify phosphorylation stoichiometry across multiple sites on intact proteins.
Phosphoprotein analysis is vital for deciphering cellular states and responses.

Purpose of the Study:

To develop and validate a novel analytical platform for comprehensive characterization of intact phosphoproteins.
To enable accurate determination of multisite phosphorylation patterns and stoichiometries.
To advance the study of post-translational modifications in complex biological systems.

Main Methods:

Integration of bottom-up and top-down proteomics approaches.
Utilizing intact protein reversed-phase liquid chromatography (RP-LC) coupled with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry.
Development of a "metal-free" RP-LC-ESI-MS platform to overcome metal ion complexation issues.

Main Results:

Demonstrated enhanced sensitivity for phosphorylated casein proteins, identifying over 20 isoforms.
Successfully applied the platform to an enriched yeast phosphoproteome, revealing multiple phosphorylation isoforms in ~16% of detected proteins.
The metal-free platform effectively characterized combinatorial post-translational modifications, particularly multisite phosphorylation.

Conclusions:

The developed intact protein LC/MS platform provides a powerful capability for comprehensive phosphoprotein analysis.
This technology significantly improves the understanding of multisite phosphorylation's role in cellular signaling.
The platform holds promise for advancing research into the complex roles of post-translational modifications in cellular regulation.