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

A new maleimide-bound acid-cleavable solid-support reagent for profiling phosphorylation.

Saiful M Chowdhury1, Gerhard R Munske, William F Siems

  • 1Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA.

Rapid Communications in Mass Spectrometry : RCM
|March 2, 2005
PubMed
Summary

This study introduces a new chemical method for phosphopeptide profiling. The technique uses solid-phase capture to selectively enrich and quantify phosphopeptides from complex biological samples.

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

Mitochondrial respiratory capacity in kidney podocytes is high, age-dependent, and sexually dimorphic.

Kidney international·2026
Same author

Site Confidence Scores among All Available Structures to Help Infer Conformational Origins of Identified Cross-Links.

Journal of proteome research·2026
Same author

On the Feasibility of Clinical Studies with Cross-Linking Mass Spectrometry.

Journal of proteome research·2026
Same author

Selective Probing of Acidic Amino Acid Residues for Protein Structural Analysis by Covalent Labeling and Mass Spectrometry.

Analytical chemistry·2025
Same author

Long AKAP18 isoforms anchor ubiquitin specific proteinases and coordinate calcium reuptake at the sarcoplasmic reticulum.

The Journal of biological chemistry·2025
Same author

Contemporary insights into elamipretide's mitochondrial mechanism of action and therapeutic effects.

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie·2025

Area of Science:

  • Biochemistry
  • Chemical Biology
  • Proteomics

Background:

  • Protein phosphorylation is a critical post-translational modification regulating cellular processes.
  • Accurate profiling of phosphopeptides is essential for understanding cell signaling and disease.
  • Existing methods for phosphopeptide enrichment can be complex and lack efficiency.

Purpose of the Study:

  • To develop a generalized and efficient chemical strategy for phosphopeptide enrichment and profiling.
  • To enable selective isolation and relative quantitation of phosphopeptides from complex biological mixtures.
  • To demonstrate the applicability of the method in real biological samples.

Main Methods:

  • A novel strategy employing solid-phase capture and mass-encoding for phosphopeptide enrichment.

Related Experiment Videos

  • Conversion of phosphates to thiols for selective isolation using a maleimide reagent.
  • Incorporation of dithiol linkers (ethanedithiol, propanedithiol) for relative quantitation via mass tagging.

    • Main Results:

    • Successful selective enrichment of phosphopeptides from model proteins (beta-casein, alpha-casein, ovalbumin).
    • Demonstration of relative quantitation using mass tagging, highlighting the need for normalization.
    • Application of the method to unfractionated whole histone protein mixtures, proving utility in complex biological samples.

    Conclusions:

    • The developed chemical strategy offers a robust method for high-throughput phosphopeptide profiling.
    • This approach simplifies reagent production and integrates with standard solid-phase peptide synthesis.
    • The method holds potential for profiling other post-translational modifications beyond phosphorylation.