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 Experiment Videos

Comparative phosphorylation site mapping from gel-derived proteins using a multidimensional ES/MS-based approach.

Francesca Zappacosta1, Michael J Huddleston, Roland S Annan

  • 1Proteomics and Biological Mass Spectrometry Laboratory, Department of Computational, Analytical and Structural Sciences, GlaxoSmithKline, King of Prussia, PA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 3, 2004
PubMed
Summary

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

A Synergistic Inhibitor Development Strategy Against Human UDP-Galactose-4-Epimerase.

Angewandte Chemie (International ed. in English)·2026
Same author

Birth rates and migration: an AI and One Health approach within the Italian National Health System.

Frontiers in public health·2026
Same author

Novel WRN Helicase Inhibitors Selectively Target Microsatellite-Unstable Cancer Cells.

Cancer discovery·2024
Same author

Reactive fragments targeting carboxylate residues employing direct to biology, high-throughput chemistry.

RSC medicinal chemistry·2023
Same author

Efficient Ligand Discovery Using Sulfur(VI) Fluoride Reactive Fragments.

ACS chemical biology·2023
Same author

Advances in high-throughput mass spectrometry in drug discovery.

EMBO molecular medicine·2022

This study introduces a novel mass spectrometry method for mapping protein phosphorylation sites. The technique helps identify and analyze multiple phosphorylation sites on proteins, crucial for understanding cell signaling pathways.

Area of Science:

  • Biochemistry
  • Proteomics
  • Molecular Biology

Background:

  • Protein phosphorylation is a key regulatory mechanism in cellular signaling pathways.
  • Identifying specific phosphorylation sites and their extent is challenging due to substoichiometric modifications.
  • Understanding protein behavior requires detailed knowledge of phosphorylation patterns.

Purpose of the Study:

  • To develop and describe a multidimensional electrospray mass spectrometry (ES-MS) based phosphopeptide-mapping strategy.
  • To provide a method for analyzing complex phosphoprotein modifications.
  • To facilitate the investigation of phosphorylation-dependent protein function.

Main Methods:

  • Selective detection and collection of phosphopeptides using on-line liquid chromatography (LC)/ES/MS.

Related Experiment Videos

  • Monitoring for phosphopeptide-specific marker ions to generate a phosphorylation profile.
  • Second-dimension analysis using nano-ES with precursor-ion scan for marker ion m/z 79.
  • Direct sequencing of phosphopeptides via LC/ES/MS/MS.

    • Main Results:

    • The developed strategy enables selective detection and fractionation of phosphopeptides.
    • A comprehensive phosphorylation profile of a protein can be generated.
    • The method allows for the identification of substoichiometric phosphorylation sites.

    Conclusions:

    • The described multidimensional ES-MS strategy is effective for phosphopeptide mapping.
    • This approach aids in assessing changes in protein phosphorylation states.
    • The technique offers a valuable tool for detailed analysis of phosphoproteins in signaling research.